bg01v  (ATCC)


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    Name:
    hESC BG01V
    Description:

    Catalog Number:
    scrc-2002
    Price:
    None
    Applications:
    BG01V cells are pluripotent and can differentiate to representatives of the three primary germ layers.
    Host:
    Homo sapiens, human
    Cell Type:
    embryonic stem cell
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    Structured Review

    ATCC bg01v
    Analysis of reverse transcriptase polymerase chain reactions (RT–PCR) of limbal stromal cells. A : Expression of SSEA-4 transcripts by limbal stromal (LS) cells as compared to human embryonic stem cells, <t>BG01V.</t> B : Expression of various transcripts by limbal stromal cells such as ABCG2, p63, Pax 6, AE5, and keratocan sulfate was compared to corneal stromal (CS) and corneal epithelial (CE) cells. GAPDH was served as housekeeping gene.

    https://www.bioz.com/result/bg01v/product/ATCC
    Average 93 stars, based on 3 article reviews
    Price from $9.99 to $1999.99
    bg01v - by Bioz Stars, 2020-07
    93/100 stars

    Images

    1) Product Images from "Ex vivo expanded SSEA-4+ human limbal stromal cells are multipotent and do not express other embryonic stem cell markers"

    Article Title: Ex vivo expanded SSEA-4+ human limbal stromal cells are multipotent and do not express other embryonic stem cell markers

    Journal: Molecular Vision

    doi:

    Analysis of reverse transcriptase polymerase chain reactions (RT–PCR) of limbal stromal cells. A : Expression of SSEA-4 transcripts by limbal stromal (LS) cells as compared to human embryonic stem cells, BG01V. B : Expression of various transcripts by limbal stromal cells such as ABCG2, p63, Pax 6, AE5, and keratocan sulfate was compared to corneal stromal (CS) and corneal epithelial (CE) cells. GAPDH was served as housekeeping gene.
    Figure Legend Snippet: Analysis of reverse transcriptase polymerase chain reactions (RT–PCR) of limbal stromal cells. A : Expression of SSEA-4 transcripts by limbal stromal (LS) cells as compared to human embryonic stem cells, BG01V. B : Expression of various transcripts by limbal stromal cells such as ABCG2, p63, Pax 6, AE5, and keratocan sulfate was compared to corneal stromal (CS) and corneal epithelial (CE) cells. GAPDH was served as housekeeping gene.

    Techniques Used: Reverse Transcription Polymerase Chain Reaction, Expressing

    2) Product Images from "Ex vivo expanded SSEA-4+ human limbal stromal cells are multipotent and do not express other embryonic stem cell markers"

    Article Title: Ex vivo expanded SSEA-4+ human limbal stromal cells are multipotent and do not express other embryonic stem cell markers

    Journal: Molecular Vision

    doi:

    Analysis of reverse transcriptase polymerase chain reactions (RT–PCR) of limbal stromal cells. A : Expression of SSEA-4 transcripts by limbal stromal (LS) cells as compared to human embryonic stem cells, BG01V. B : Expression of various transcripts by limbal stromal cells such as ABCG2, p63, Pax 6, AE5, and keratocan sulfate was compared to corneal stromal (CS) and corneal epithelial (CE) cells. GAPDH was served as housekeeping gene.
    Figure Legend Snippet: Analysis of reverse transcriptase polymerase chain reactions (RT–PCR) of limbal stromal cells. A : Expression of SSEA-4 transcripts by limbal stromal (LS) cells as compared to human embryonic stem cells, BG01V. B : Expression of various transcripts by limbal stromal cells such as ABCG2, p63, Pax 6, AE5, and keratocan sulfate was compared to corneal stromal (CS) and corneal epithelial (CE) cells. GAPDH was served as housekeeping gene.

    Techniques Used: Reverse Transcription Polymerase Chain Reaction, Expressing

    3) Product Images from "Astrocytes derived from trisomic human embryonic stem cells express markers of astrocytic cancer cells and premalignant stem-like progenitors"

    Article Title: Astrocytes derived from trisomic human embryonic stem cells express markers of astrocytic cancer cells and premalignant stem-like progenitors

    Journal: BMC Medical Genomics

    doi: 10.1186/1755-8794-3-12

    Gene expression profile of trisomic BG01V APCs is similar to glioblastomas . A. Heat map of unsupervised hierarchical cluster analysis of microarray hybridization data derived from the twenty-three glioblastoma patient samples (N), the three trisomic BG01V APC samples (G) and the three diploid H9 APC samples (C). Relative over expression is indicated by red and under expression by blue in the heat map. Data filtered using a p value of
    Figure Legend Snippet: Gene expression profile of trisomic BG01V APCs is similar to glioblastomas . A. Heat map of unsupervised hierarchical cluster analysis of microarray hybridization data derived from the twenty-three glioblastoma patient samples (N), the three trisomic BG01V APC samples (G) and the three diploid H9 APC samples (C). Relative over expression is indicated by red and under expression by blue in the heat map. Data filtered using a p value of

    Techniques Used: Expressing, Microarray, Hybridization, Derivative Assay, Over Expression

    Over expressed transcripts include known and novel markers of premalignant astrocytic stem/progenitor cells . Individual gene dot plots showing expression levels of several over expressed transcripts in all twenty-three glioblastoma patient samples (purple, N), three trisomic BG01V APCs samples (green, G), three CCF-STTG1 astrocytoma cell samples (blue, D) and diploid H9 APCs (red, C). Transcripts shown are PROM1 (panel A), CHI3L1 (panel B), RGS5 (panel C), IGFBP2 (panel D), PPP2R2B (panel E), LPHN2 (panel F), KCNMB4 (panel G), ASTN1 (panel H) and GPC4 (panel I).
    Figure Legend Snippet: Over expressed transcripts include known and novel markers of premalignant astrocytic stem/progenitor cells . Individual gene dot plots showing expression levels of several over expressed transcripts in all twenty-three glioblastoma patient samples (purple, N), three trisomic BG01V APCs samples (green, G), three CCF-STTG1 astrocytoma cell samples (blue, D) and diploid H9 APCs (red, C). Transcripts shown are PROM1 (panel A), CHI3L1 (panel B), RGS5 (panel C), IGFBP2 (panel D), PPP2R2B (panel E), LPHN2 (panel F), KCNMB4 (panel G), ASTN1 (panel H) and GPC4 (panel I).

    Techniques Used: Expressing

    Intersection of microarray data identifies markers of premalignant astrocytic stem-like/progenitor cells . Venn diagram depicting the overlap between the four pair wise comparisons: GvC, GvD, NvC and NvD that was used to identify the 311 transcripts exhibiting similar changes in expression patterns in trisomic BG01V APCs (samples G) and glioblastoma patients (samples N) relative to diploid H9 APCs (samples C) and CCF-STTG1 astrocytoma cells (samples D), which constitutes the GNvCD gene list shown in Additional file 6 , Table S5.
    Figure Legend Snippet: Intersection of microarray data identifies markers of premalignant astrocytic stem-like/progenitor cells . Venn diagram depicting the overlap between the four pair wise comparisons: GvC, GvD, NvC and NvD that was used to identify the 311 transcripts exhibiting similar changes in expression patterns in trisomic BG01V APCs (samples G) and glioblastoma patients (samples N) relative to diploid H9 APCs (samples C) and CCF-STTG1 astrocytoma cells (samples D), which constitutes the GNvCD gene list shown in Additional file 6 , Table S5.

    Techniques Used: Microarray, Expressing

    Global gene expression profile of trisomic BG01V APCs is similar to CCF-STTG1 astrocytoma cells . A. Heat map of unsupervised hierarchical cluster analysis of exon microarray hybridization data derived from the three diploid H9 APC samples (C), the three trisomic BG01V APC samples (G) and the three CCF-STTG1 astrocytoma cell samples (D). Relative over expression is indicated by red and under expression by blue in the heat map. Microarray data was filtered using a p value
    Figure Legend Snippet: Global gene expression profile of trisomic BG01V APCs is similar to CCF-STTG1 astrocytoma cells . A. Heat map of unsupervised hierarchical cluster analysis of exon microarray hybridization data derived from the three diploid H9 APC samples (C), the three trisomic BG01V APC samples (G) and the three CCF-STTG1 astrocytoma cell samples (D). Relative over expression is indicated by red and under expression by blue in the heat map. Microarray data was filtered using a p value

    Techniques Used: Expressing, Microarray, Hybridization, Derivative Assay, Over Expression

    Intersection of microarray data identifies common markers of astrocytic cancer cells . Venn diagram depicting the overlap, identified by the supervised hierarchical analysis, between the two pair wise comparisons, GvC and DvC, that was used to identify the 1038 transcripts exhibiting consistent sign changes in expression patterns in trisomic BG01V APCs (samples G) and the CCF-STTG1 astrocytoma cell line (samples D) relative to diploid H9 APCs (samples C), which constitutes the GDvC gene list shown in Additional file 2 , Table S2.
    Figure Legend Snippet: Intersection of microarray data identifies common markers of astrocytic cancer cells . Venn diagram depicting the overlap, identified by the supervised hierarchical analysis, between the two pair wise comparisons, GvC and DvC, that was used to identify the 1038 transcripts exhibiting consistent sign changes in expression patterns in trisomic BG01V APCs (samples G) and the CCF-STTG1 astrocytoma cell line (samples D) relative to diploid H9 APCs (samples C), which constitutes the GDvC gene list shown in Additional file 2 , Table S2.

    Techniques Used: Microarray, Expressing

    Similarities in expression changes in trisomic APCs and astrocytoma cells were validated by RT-PCR analyses . A. Semi-quantitative RT-PCR analysis shows relative under expression of COL4A6 , GABRA2 , MGMT and TRPA1 (left panels), and over expression of HDAC9 , CPXM1 , DNMT3A, GUCY1A3 and STAT3 (right panels) transcripts in trisomic BG01V APCs and CCF-STTG1 astrocytoma cells with respect to diploid H9 APCs. 18S RNA was used as a control. B. Quantitative RT-PCR validation of relative changes in expression levels of the transcripts shown in panel A in diploid H9 APCs (blue bars), trisomic BG01V APCs (yellow bars) and astrocytoma cells (red bars).
    Figure Legend Snippet: Similarities in expression changes in trisomic APCs and astrocytoma cells were validated by RT-PCR analyses . A. Semi-quantitative RT-PCR analysis shows relative under expression of COL4A6 , GABRA2 , MGMT and TRPA1 (left panels), and over expression of HDAC9 , CPXM1 , DNMT3A, GUCY1A3 and STAT3 (right panels) transcripts in trisomic BG01V APCs and CCF-STTG1 astrocytoma cells with respect to diploid H9 APCs. 18S RNA was used as a control. B. Quantitative RT-PCR validation of relative changes in expression levels of the transcripts shown in panel A in diploid H9 APCs (blue bars), trisomic BG01V APCs (yellow bars) and astrocytoma cells (red bars).

    Techniques Used: Expressing, Reverse Transcription Polymerase Chain Reaction, Quantitative RT-PCR, Over Expression

    Intersection of microarray data identifies markers of astrocytic cancer cells . Venn diagram depicting the overlap between the two datasets, GDvC (Additional file 2 , Table S2) and GNvC (Additional file 4 , Table S3) that was used to identify the 499 transcripts exhibiting similar changes in expression patterns in all aneuploid astrocytic cell populations including trisomic BG01V APCs (samples G), glioblastoma patients (samples N) and CCF-STTG1 astrocytoma cells (samples D) relative to normal, diploid H9-derived APCs (samples C), which constitutes the GNDvC gene list shown in Additional file 5 , Table S4.
    Figure Legend Snippet: Intersection of microarray data identifies markers of astrocytic cancer cells . Venn diagram depicting the overlap between the two datasets, GDvC (Additional file 2 , Table S2) and GNvC (Additional file 4 , Table S3) that was used to identify the 499 transcripts exhibiting similar changes in expression patterns in all aneuploid astrocytic cell populations including trisomic BG01V APCs (samples G), glioblastoma patients (samples N) and CCF-STTG1 astrocytoma cells (samples D) relative to normal, diploid H9-derived APCs (samples C), which constitutes the GNDvC gene list shown in Additional file 5 , Table S4.

    Techniques Used: Microarray, Expressing, Derivative Assay

    Diploid and trisomic astrocytic progenitors derived by directed in vitro differentiation of hESCs exhibit transcript and alternative splicing patterns characteristic of astrocytic cells . A. Diploid H9 and trisomic BG01V hESCs colonies (unstained, left panels), express stem cell markers OCT-4 (red) and TRA-1-60 (green). Karyotypes of diploid H9 (46; XX) and trisomic BG01V (49; XXY, +12, +17) hESCs. Diploid and trisomic APCs express astrocytic marker, GFAP (red). B. Semiquantitative RT-PCR analysis demonstrating that stem cell transcripts Noggin (top panel) and Lin28 (second panel) are over expressed in both diploid H9 and trisomic BG01V hESC lines relative to all astrocytic-like cells including H9 APCs, BG01V APCs and CCF-STTG1 astrocytoma cells. GFAP transcripts are up regulated in astrocytic cells relative to hESCs (middle panel). Alternative splicing pattern of FGFR1 in diploid H9 hESCs is preserved in trisomic BG01V hESCs relative to all astrocytic cells. 18S RNA was used as control in all RT-PCR experiments (bottom panel). MW marker is included in the far left lane in each gel. C. DNA sequence analysis demonstrating that the FGFR1 transcript expressed in diploid H9 and trisomic BG01V hESCs includes exon 3, while astrocytic cells express a FGFR1 splice variant in which exon 3 has been excluded (from FGFR1 RT-PCR analysis in Figure 1B).
    Figure Legend Snippet: Diploid and trisomic astrocytic progenitors derived by directed in vitro differentiation of hESCs exhibit transcript and alternative splicing patterns characteristic of astrocytic cells . A. Diploid H9 and trisomic BG01V hESCs colonies (unstained, left panels), express stem cell markers OCT-4 (red) and TRA-1-60 (green). Karyotypes of diploid H9 (46; XX) and trisomic BG01V (49; XXY, +12, +17) hESCs. Diploid and trisomic APCs express astrocytic marker, GFAP (red). B. Semiquantitative RT-PCR analysis demonstrating that stem cell transcripts Noggin (top panel) and Lin28 (second panel) are over expressed in both diploid H9 and trisomic BG01V hESC lines relative to all astrocytic-like cells including H9 APCs, BG01V APCs and CCF-STTG1 astrocytoma cells. GFAP transcripts are up regulated in astrocytic cells relative to hESCs (middle panel). Alternative splicing pattern of FGFR1 in diploid H9 hESCs is preserved in trisomic BG01V hESCs relative to all astrocytic cells. 18S RNA was used as control in all RT-PCR experiments (bottom panel). MW marker is included in the far left lane in each gel. C. DNA sequence analysis demonstrating that the FGFR1 transcript expressed in diploid H9 and trisomic BG01V hESCs includes exon 3, while astrocytic cells express a FGFR1 splice variant in which exon 3 has been excluded (from FGFR1 RT-PCR analysis in Figure 1B).

    Techniques Used: Derivative Assay, In Vitro, Marker, Reverse Transcription Polymerase Chain Reaction, Sequencing, Variant Assay

    4) Product Images from "Ex vivo expanded SSEA-4+ human limbal stromal cells are multipotent and do not express other embryonic stem cell markers"

    Article Title: Ex vivo expanded SSEA-4+ human limbal stromal cells are multipotent and do not express other embryonic stem cell markers

    Journal: Molecular Vision

    doi:

    Analysis of reverse transcriptase polymerase chain reactions (RT–PCR) of limbal stromal cells. A : Expression of SSEA-4 transcripts by limbal stromal (LS) cells as compared to human embryonic stem cells, BG01V. B : Expression of various transcripts by limbal stromal cells such as ABCG2, p63, Pax 6, AE5, and keratocan sulfate was compared to corneal stromal (CS) and corneal epithelial (CE) cells. GAPDH was served as housekeeping gene.
    Figure Legend Snippet: Analysis of reverse transcriptase polymerase chain reactions (RT–PCR) of limbal stromal cells. A : Expression of SSEA-4 transcripts by limbal stromal (LS) cells as compared to human embryonic stem cells, BG01V. B : Expression of various transcripts by limbal stromal cells such as ABCG2, p63, Pax 6, AE5, and keratocan sulfate was compared to corneal stromal (CS) and corneal epithelial (CE) cells. GAPDH was served as housekeeping gene.

    Techniques Used: Reverse Transcription Polymerase Chain Reaction, Expressing

    5) Product Images from "Astrocytes derived from trisomic human embryonic stem cells express markers of astrocytic cancer cells and premalignant stem-like progenitors"

    Article Title: Astrocytes derived from trisomic human embryonic stem cells express markers of astrocytic cancer cells and premalignant stem-like progenitors

    Journal: BMC Medical Genomics

    doi: 10.1186/1755-8794-3-12

    Gene expression profile of trisomic BG01V APCs is similar to glioblastomas . A. Heat map of unsupervised hierarchical cluster analysis of microarray hybridization data derived from the twenty-three glioblastoma patient samples (N), the three trisomic BG01V APC samples (G) and the three diploid H9 APC samples (C). Relative over expression is indicated by red and under expression by blue in the heat map. Data filtered using a p value of
    Figure Legend Snippet: Gene expression profile of trisomic BG01V APCs is similar to glioblastomas . A. Heat map of unsupervised hierarchical cluster analysis of microarray hybridization data derived from the twenty-three glioblastoma patient samples (N), the three trisomic BG01V APC samples (G) and the three diploid H9 APC samples (C). Relative over expression is indicated by red and under expression by blue in the heat map. Data filtered using a p value of

    Techniques Used: Expressing, Microarray, Hybridization, Derivative Assay, Over Expression

    Over expressed transcripts include known and novel markers of premalignant astrocytic stem/progenitor cells . Individual gene dot plots showing expression levels of several over expressed transcripts in all twenty-three glioblastoma patient samples (purple, N), three trisomic BG01V APCs samples (green, G), three CCF-STTG1 astrocytoma cell samples (blue, D) and diploid H9 APCs (red, C). Transcripts shown are PROM1 (panel A), CHI3L1 (panel B), RGS5 (panel C), IGFBP2 (panel D), PPP2R2B (panel E), LPHN2 (panel F), KCNMB4 (panel G), ASTN1 (panel H) and GPC4 (panel I).
    Figure Legend Snippet: Over expressed transcripts include known and novel markers of premalignant astrocytic stem/progenitor cells . Individual gene dot plots showing expression levels of several over expressed transcripts in all twenty-three glioblastoma patient samples (purple, N), three trisomic BG01V APCs samples (green, G), three CCF-STTG1 astrocytoma cell samples (blue, D) and diploid H9 APCs (red, C). Transcripts shown are PROM1 (panel A), CHI3L1 (panel B), RGS5 (panel C), IGFBP2 (panel D), PPP2R2B (panel E), LPHN2 (panel F), KCNMB4 (panel G), ASTN1 (panel H) and GPC4 (panel I).

    Techniques Used: Expressing

    Intersection of microarray data identifies markers of premalignant astrocytic stem-like/progenitor cells . Venn diagram depicting the overlap between the four pair wise comparisons: GvC, GvD, NvC and NvD that was used to identify the 311 transcripts exhibiting similar changes in expression patterns in trisomic BG01V APCs (samples G) and glioblastoma patients (samples N) relative to diploid H9 APCs (samples C) and CCF-STTG1 astrocytoma cells (samples D), which constitutes the GNvCD gene list shown in Additional file 6 , Table S5.
    Figure Legend Snippet: Intersection of microarray data identifies markers of premalignant astrocytic stem-like/progenitor cells . Venn diagram depicting the overlap between the four pair wise comparisons: GvC, GvD, NvC and NvD that was used to identify the 311 transcripts exhibiting similar changes in expression patterns in trisomic BG01V APCs (samples G) and glioblastoma patients (samples N) relative to diploid H9 APCs (samples C) and CCF-STTG1 astrocytoma cells (samples D), which constitutes the GNvCD gene list shown in Additional file 6 , Table S5.

    Techniques Used: Microarray, Expressing

    Global gene expression profile of trisomic BG01V APCs is similar to CCF-STTG1 astrocytoma cells . A. Heat map of unsupervised hierarchical cluster analysis of exon microarray hybridization data derived from the three diploid H9 APC samples (C), the three trisomic BG01V APC samples (G) and the three CCF-STTG1 astrocytoma cell samples (D). Relative over expression is indicated by red and under expression by blue in the heat map. Microarray data was filtered using a p value
    Figure Legend Snippet: Global gene expression profile of trisomic BG01V APCs is similar to CCF-STTG1 astrocytoma cells . A. Heat map of unsupervised hierarchical cluster analysis of exon microarray hybridization data derived from the three diploid H9 APC samples (C), the three trisomic BG01V APC samples (G) and the three CCF-STTG1 astrocytoma cell samples (D). Relative over expression is indicated by red and under expression by blue in the heat map. Microarray data was filtered using a p value

    Techniques Used: Expressing, Microarray, Hybridization, Derivative Assay, Over Expression

    Intersection of microarray data identifies common markers of astrocytic cancer cells . Venn diagram depicting the overlap, identified by the supervised hierarchical analysis, between the two pair wise comparisons, GvC and DvC, that was used to identify the 1038 transcripts exhibiting consistent sign changes in expression patterns in trisomic BG01V APCs (samples G) and the CCF-STTG1 astrocytoma cell line (samples D) relative to diploid H9 APCs (samples C), which constitutes the GDvC gene list shown in Additional file 2 , Table S2.
    Figure Legend Snippet: Intersection of microarray data identifies common markers of astrocytic cancer cells . Venn diagram depicting the overlap, identified by the supervised hierarchical analysis, between the two pair wise comparisons, GvC and DvC, that was used to identify the 1038 transcripts exhibiting consistent sign changes in expression patterns in trisomic BG01V APCs (samples G) and the CCF-STTG1 astrocytoma cell line (samples D) relative to diploid H9 APCs (samples C), which constitutes the GDvC gene list shown in Additional file 2 , Table S2.

    Techniques Used: Microarray, Expressing

    Similarities in expression changes in trisomic APCs and astrocytoma cells were validated by RT-PCR analyses . A. Semi-quantitative RT-PCR analysis shows relative under expression of COL4A6 , GABRA2 , MGMT and TRPA1 (left panels), and over expression of HDAC9 , CPXM1 , DNMT3A, GUCY1A3 and STAT3 (right panels) transcripts in trisomic BG01V APCs and CCF-STTG1 astrocytoma cells with respect to diploid H9 APCs. 18S RNA was used as a control. B. Quantitative RT-PCR validation of relative changes in expression levels of the transcripts shown in panel A in diploid H9 APCs (blue bars), trisomic BG01V APCs (yellow bars) and astrocytoma cells (red bars).
    Figure Legend Snippet: Similarities in expression changes in trisomic APCs and astrocytoma cells were validated by RT-PCR analyses . A. Semi-quantitative RT-PCR analysis shows relative under expression of COL4A6 , GABRA2 , MGMT and TRPA1 (left panels), and over expression of HDAC9 , CPXM1 , DNMT3A, GUCY1A3 and STAT3 (right panels) transcripts in trisomic BG01V APCs and CCF-STTG1 astrocytoma cells with respect to diploid H9 APCs. 18S RNA was used as a control. B. Quantitative RT-PCR validation of relative changes in expression levels of the transcripts shown in panel A in diploid H9 APCs (blue bars), trisomic BG01V APCs (yellow bars) and astrocytoma cells (red bars).

    Techniques Used: Expressing, Reverse Transcription Polymerase Chain Reaction, Quantitative RT-PCR, Over Expression

    Intersection of microarray data identifies markers of astrocytic cancer cells . Venn diagram depicting the overlap between the two datasets, GDvC (Additional file 2 , Table S2) and GNvC (Additional file 4 , Table S3) that was used to identify the 499 transcripts exhibiting similar changes in expression patterns in all aneuploid astrocytic cell populations including trisomic BG01V APCs (samples G), glioblastoma patients (samples N) and CCF-STTG1 astrocytoma cells (samples D) relative to normal, diploid H9-derived APCs (samples C), which constitutes the GNDvC gene list shown in Additional file 5 , Table S4.
    Figure Legend Snippet: Intersection of microarray data identifies markers of astrocytic cancer cells . Venn diagram depicting the overlap between the two datasets, GDvC (Additional file 2 , Table S2) and GNvC (Additional file 4 , Table S3) that was used to identify the 499 transcripts exhibiting similar changes in expression patterns in all aneuploid astrocytic cell populations including trisomic BG01V APCs (samples G), glioblastoma patients (samples N) and CCF-STTG1 astrocytoma cells (samples D) relative to normal, diploid H9-derived APCs (samples C), which constitutes the GNDvC gene list shown in Additional file 5 , Table S4.

    Techniques Used: Microarray, Expressing, Derivative Assay

    Diploid and trisomic astrocytic progenitors derived by directed in vitro differentiation of hESCs exhibit transcript and alternative splicing patterns characteristic of astrocytic cells . A. Diploid H9 and trisomic BG01V hESCs colonies (unstained, left panels), express stem cell markers OCT-4 (red) and TRA-1-60 (green). Karyotypes of diploid H9 (46; XX) and trisomic BG01V (49; XXY, +12, +17) hESCs. Diploid and trisomic APCs express astrocytic marker, GFAP (red). B. Semiquantitative RT-PCR analysis demonstrating that stem cell transcripts Noggin (top panel) and Lin28 (second panel) are over expressed in both diploid H9 and trisomic BG01V hESC lines relative to all astrocytic-like cells including H9 APCs, BG01V APCs and CCF-STTG1 astrocytoma cells. GFAP transcripts are up regulated in astrocytic cells relative to hESCs (middle panel). Alternative splicing pattern of FGFR1 in diploid H9 hESCs is preserved in trisomic BG01V hESCs relative to all astrocytic cells. 18S RNA was used as control in all RT-PCR experiments (bottom panel). MW marker is included in the far left lane in each gel. C. DNA sequence analysis demonstrating that the FGFR1 transcript expressed in diploid H9 and trisomic BG01V hESCs includes exon 3, while astrocytic cells express a FGFR1 splice variant in which exon 3 has been excluded (from FGFR1 RT-PCR analysis in Figure 1B).
    Figure Legend Snippet: Diploid and trisomic astrocytic progenitors derived by directed in vitro differentiation of hESCs exhibit transcript and alternative splicing patterns characteristic of astrocytic cells . A. Diploid H9 and trisomic BG01V hESCs colonies (unstained, left panels), express stem cell markers OCT-4 (red) and TRA-1-60 (green). Karyotypes of diploid H9 (46; XX) and trisomic BG01V (49; XXY, +12, +17) hESCs. Diploid and trisomic APCs express astrocytic marker, GFAP (red). B. Semiquantitative RT-PCR analysis demonstrating that stem cell transcripts Noggin (top panel) and Lin28 (second panel) are over expressed in both diploid H9 and trisomic BG01V hESC lines relative to all astrocytic-like cells including H9 APCs, BG01V APCs and CCF-STTG1 astrocytoma cells. GFAP transcripts are up regulated in astrocytic cells relative to hESCs (middle panel). Alternative splicing pattern of FGFR1 in diploid H9 hESCs is preserved in trisomic BG01V hESCs relative to all astrocytic cells. 18S RNA was used as control in all RT-PCR experiments (bottom panel). MW marker is included in the far left lane in each gel. C. DNA sequence analysis demonstrating that the FGFR1 transcript expressed in diploid H9 and trisomic BG01V hESCs includes exon 3, while astrocytic cells express a FGFR1 splice variant in which exon 3 has been excluded (from FGFR1 RT-PCR analysis in Figure 1B).

    Techniques Used: Derivative Assay, In Vitro, Marker, Reverse Transcription Polymerase Chain Reaction, Sequencing, Variant Assay

    Related Articles

    Cell Culture:

    Article Title: Ex vivo expanded SSEA-4+ human limbal stromal cells are multipotent and do not express other embryonic stem cell markers
    Article Snippet: .. Human embryonic stem cell (hESC) culture Embryonic stem cell line, BG01V (ATCC) at passage 16 were cultured on mitomycin treated MEF cells (ATCC) with 80% knockout DMEM supplemented with 20% Gibco knockout SR, 1% MEM-non essential amino acid (NEAA), 1% Glutamax, 0.1 mM B-mercaptoethanol (BME), 10 IU/ml penicillin, 10 µg/ml streptomycin (all from Invitrogen Corporation), and 4 ng/ml human basic fibroblast growth factor (bFGF; BD Biosciences) [ ]. .. Cell colonies at 70% confluence were harvested with collagenase and gently scrapped with 5 ml serological pipette.

    Article Title: Effect of cellular mass on chondrogenic differentiation during embryoid body formation
    Article Snippet: .. HESC culture The BG01V hESC line (American Type Culture Collection, Manassas, VA; USA) was cultured in mitomycin-C-treated mouse embryonic fibroblasts (MEFs; passage 3; Sigma-Aldrich; Merck KGaA, Darmstadt, Germany) seeded on a culture dish previously coated with Matrigel™ (Corning Incorporated, Corning, NY, USA). .. The hESCs were cultured in DMEM/F12 (Merck Millipore, Germany) supplemented with 20% KnockOut™ Serum Replacement (KnockOut™ SR, Thermo Fisher Scientific Inc., MA, USA), 1% penicillin-streptomycin (Sigma-Aldrich; Merck KGaA), 1 mM non-essential amino acid (NEAA; Sigma-Aldrich; Merck KGaA), 0.2 mM 2-mercaptoethanol (Sigma-Aldrich; Merck KGaA), and 10 ng/ml basic fibroblast growth factor (bFGF) (Sigma-Aldrich; Merck KGaA).

    Article Title: Astrocytes derived from trisomic human embryonic stem cells express markers of astrocytic cancer cells and premalignant stem-like progenitors
    Article Snippet: .. HESC and other cell culture HESC lines H9 (WiCell) and BG01V (American Type Culture Collection) were grown under feeder-independent conditions on matrigel-coated dishes (BD) in medium containing basal DMEM/F-12 with 1 mM glutamine, 20% knockout serum replacement (KSR; Invitrogen), 2 mM non-essential amino acids and 8 ng/ml FGF (Invitrogen). .. To obtain non-adherent embryoid bodies, small pieces of undifferentiated hESC colonies (~100-200 cells) were mechanically dissected and cultured on low attachment plates in the same media used for maintaining pluripotent hESCs, except KSR was removed and replaced with 10% Fetal Bovine Serum (FBS; Hyclone).

    Knock-Out:

    Article Title: Ex vivo expanded SSEA-4+ human limbal stromal cells are multipotent and do not express other embryonic stem cell markers
    Article Snippet: .. Human embryonic stem cell (hESC) culture Embryonic stem cell line, BG01V (ATCC) at passage 16 were cultured on mitomycin treated MEF cells (ATCC) with 80% knockout DMEM supplemented with 20% Gibco knockout SR, 1% MEM-non essential amino acid (NEAA), 1% Glutamax, 0.1 mM B-mercaptoethanol (BME), 10 IU/ml penicillin, 10 µg/ml streptomycin (all from Invitrogen Corporation), and 4 ng/ml human basic fibroblast growth factor (bFGF; BD Biosciences) [ ]. .. Cell colonies at 70% confluence were harvested with collagenase and gently scrapped with 5 ml serological pipette.

    Article Title: SOX2 and OCT4 mRNA-Expressing Cells, Detected by Molecular Beacons, Localize to the Center of Neurospheres during Differentiation
    Article Snippet: .. The human embryonic stem cell (hESC) line BG01V (ATCC, SCRC-2002) was propagated on Mitomycin C-treated mouse embryonic fibroblasts (ATCC, MEF SCRC-1040) in complete GM DMEM/F12 containing 2mM L-glutamine, 1x MEM non-essential amino acid solution (Gibco), 0.1mM 2-mercaptoethanol (Sigma), 4ng/ml bFGF (Invitrogen), 5% knockout serum replacement (Invitrogen), 15% fetal bovine serum (FBS), and 1% penicillin/streptomycin. ..

    Article Title: Astrocytes derived from trisomic human embryonic stem cells express markers of astrocytic cancer cells and premalignant stem-like progenitors
    Article Snippet: .. HESC and other cell culture HESC lines H9 (WiCell) and BG01V (American Type Culture Collection) were grown under feeder-independent conditions on matrigel-coated dishes (BD) in medium containing basal DMEM/F-12 with 1 mM glutamine, 20% knockout serum replacement (KSR; Invitrogen), 2 mM non-essential amino acids and 8 ng/ml FGF (Invitrogen). .. To obtain non-adherent embryoid bodies, small pieces of undifferentiated hESC colonies (~100-200 cells) were mechanically dissected and cultured on low attachment plates in the same media used for maintaining pluripotent hESCs, except KSR was removed and replaced with 10% Fetal Bovine Serum (FBS; Hyclone).

    Expressing:

    Article Title: Ex vivo expanded SSEA-4+ human limbal stromal cells are multipotent and do not express other embryonic stem cell markers
    Article Snippet: .. Reverse transcription polymerase chain reaction (RT–PCR) analysis Expression of embryonic protein SSEA-4 by limbal stromal cells was compared to hESC, BG01V. ..

    Article Title: Ex vivo expanded SSEA-4+ human limbal stromal cells are multipotent and do not express other embryonic stem cell markers
    Article Snippet: .. RT–PCR analysis Our results confirmed the expression of SSEA-4 in the limbal stromal cells as compared to hESC, BG01V ( ). ..

    Reverse Transcription Polymerase Chain Reaction:

    Article Title: Ex vivo expanded SSEA-4+ human limbal stromal cells are multipotent and do not express other embryonic stem cell markers
    Article Snippet: .. Reverse transcription polymerase chain reaction (RT–PCR) analysis Expression of embryonic protein SSEA-4 by limbal stromal cells was compared to hESC, BG01V. ..

    Article Title: Ex vivo expanded SSEA-4+ human limbal stromal cells are multipotent and do not express other embryonic stem cell markers
    Article Snippet: .. RT–PCR analysis Our results confirmed the expression of SSEA-4 in the limbal stromal cells as compared to hESC, BG01V ( ). ..

    Transformation Assay:

    Article Title: Astrocytes derived from trisomic human embryonic stem cells express markers of astrocytic cancer cells and premalignant stem-like progenitors
    Article Snippet: .. However, we cannot rule out the possibility that genetic events other than trisomy played a role in the initiation of premalignant transformation observed here since the trisomic hESC line, BG01V, is not a derivative of the diploid hESC line, H9. .. That a constellation of gain-of-function and/or loss-of-function mutations in multiple genes is required for malignant transformation has been known for decades [ , ].

    Variant Assay:

    Article Title: Genome wide profiling of human embryonic stem cells (hESCs), their derivatives and embryonal carcinoma cells to develop base profiles of U.S. Federal government approved hESC lines
    Article Snippet: .. Samples included 7 hESC lines BG01, BG02, BG03, I6, H1, H7 and H9, EBs that were differentiated from hESCs of the three BG lines, human fibroblast feeder HS27 (ATCC), hESC-derived fibroblasts, karyotypically abnormal hESC line BG01 Variant (BG01V) [ ] and EC line NTera2 [ ]. ..

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    ATCC bg01v
    Analysis of reverse transcriptase polymerase chain reactions (RT–PCR) of limbal stromal cells. A : Expression of SSEA-4 transcripts by limbal stromal (LS) cells as compared to human embryonic stem cells, <t>BG01V.</t> B : Expression of various transcripts by limbal stromal cells such as ABCG2, p63, Pax 6, AE5, and keratocan sulfate was compared to corneal stromal (CS) and corneal epithelial (CE) cells. GAPDH was served as housekeeping gene.
    Bg01v, supplied by ATCC, used in various techniques. Bioz Stars score: 93/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    ATCC human bg01v embryonic stem cells
    <t>BG01V</t> embryonic stem cells differentiate into neural progenitors (A) Contrast phase microscopic image of ESC grown on a MEF feeder layer (left), and generated NP grown on laminin (right). Arrows point to ESC colonies. (B) Cell lysates were prepared, and the expression of the indicated markers was analyzed by Western blotting as described in the M M section. β-tubulin was used as a loading control. (C) Immunocytochemical staining of stem cells and differentiated neural progenitors with antibodies (Ab) against the cell specific markers (red/green) as indicated, and DAPI staining (blue) was performed as described in the M M section. Differentiation experiments were repeated six times with the consistent results.
    Human Bg01v Embryonic Stem Cells, supplied by ATCC, used in various techniques. Bioz Stars score: 85/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Analysis of reverse transcriptase polymerase chain reactions (RT–PCR) of limbal stromal cells. A : Expression of SSEA-4 transcripts by limbal stromal (LS) cells as compared to human embryonic stem cells, BG01V. B : Expression of various transcripts by limbal stromal cells such as ABCG2, p63, Pax 6, AE5, and keratocan sulfate was compared to corneal stromal (CS) and corneal epithelial (CE) cells. GAPDH was served as housekeeping gene.

    Journal: Molecular Vision

    Article Title: Ex vivo expanded SSEA-4+ human limbal stromal cells are multipotent and do not express other embryonic stem cell markers

    doi:

    Figure Lengend Snippet: Analysis of reverse transcriptase polymerase chain reactions (RT–PCR) of limbal stromal cells. A : Expression of SSEA-4 transcripts by limbal stromal (LS) cells as compared to human embryonic stem cells, BG01V. B : Expression of various transcripts by limbal stromal cells such as ABCG2, p63, Pax 6, AE5, and keratocan sulfate was compared to corneal stromal (CS) and corneal epithelial (CE) cells. GAPDH was served as housekeeping gene.

    Article Snippet: Human embryonic stem cell (hESC) culture Embryonic stem cell line, BG01V (ATCC) at passage 16 were cultured on mitomycin treated MEF cells (ATCC) with 80% knockout DMEM supplemented with 20% Gibco knockout SR, 1% MEM-non essential amino acid (NEAA), 1% Glutamax, 0.1 mM B-mercaptoethanol (BME), 10 IU/ml penicillin, 10 µg/ml streptomycin (all from Invitrogen Corporation), and 4 ng/ml human basic fibroblast growth factor (bFGF; BD Biosciences) [ ].

    Techniques: Reverse Transcription Polymerase Chain Reaction, Expressing

    Male and female hESC lines can be distinguished by genes identified by bead array. Five potential genes RPS4Y, RPS4Y2, EIF1AY, VCY, and AMELY are located in the Y chromosome. By comparing the expression level of these genes in all hESC lines, we have found that 3 out of 5 were specifically expressed in male hESC lines I6, BG01 and BG02 (A) and this was verified by RT-PCR in male line BG01 and female line BG03 (B). G3PDH was used as an internal control. *: represents the gene expression level is detected at

    Journal: BMC Developmental Biology

    Article Title: Genome wide profiling of human embryonic stem cells (hESCs), their derivatives and embryonal carcinoma cells to develop base profiles of U.S. Federal government approved hESC lines

    doi: 10.1186/1471-213X-6-20

    Figure Lengend Snippet: Male and female hESC lines can be distinguished by genes identified by bead array. Five potential genes RPS4Y, RPS4Y2, EIF1AY, VCY, and AMELY are located in the Y chromosome. By comparing the expression level of these genes in all hESC lines, we have found that 3 out of 5 were specifically expressed in male hESC lines I6, BG01 and BG02 (A) and this was verified by RT-PCR in male line BG01 and female line BG03 (B). G3PDH was used as an internal control. *: represents the gene expression level is detected at

    Article Snippet: Samples included 7 hESC lines BG01, BG02, BG03, I6, H1, H7 and H9, EBs that were differentiated from hESCs of the three BG lines, human fibroblast feeder HS27 (ATCC), hESC-derived fibroblasts, karyotypically abnormal hESC line BG01 Variant (BG01V) [ ] and EC line NTera2 [ ].

    Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction

    BG lines show small but distinct differences as assessed by bead array. These three hESC lines share high similarities as shown by the scatterplots of BG01 vs BG02 (B), BG01 vs BG03 (D) and BG02 vs BG03 (F). Comparisons of all three lines were made and lists of selected genes that were specifically expressed in BG01 (A), BG02 (C) and BG03 (F) are shown. Correlation coefficients (R 2 ) were generated using all genes with expression level > 0 (black and blue dots), or all genes with detection confidence > 0.99 (blue dots). Genes outside the two thin red lines were detected at > 2.5- fold difference.

    Journal: BMC Developmental Biology

    Article Title: Genome wide profiling of human embryonic stem cells (hESCs), their derivatives and embryonal carcinoma cells to develop base profiles of U.S. Federal government approved hESC lines

    doi: 10.1186/1471-213X-6-20

    Figure Lengend Snippet: BG lines show small but distinct differences as assessed by bead array. These three hESC lines share high similarities as shown by the scatterplots of BG01 vs BG02 (B), BG01 vs BG03 (D) and BG02 vs BG03 (F). Comparisons of all three lines were made and lists of selected genes that were specifically expressed in BG01 (A), BG02 (C) and BG03 (F) are shown. Correlation coefficients (R 2 ) were generated using all genes with expression level > 0 (black and blue dots), or all genes with detection confidence > 0.99 (blue dots). Genes outside the two thin red lines were detected at > 2.5- fold difference.

    Article Snippet: Samples included 7 hESC lines BG01, BG02, BG03, I6, H1, H7 and H9, EBs that were differentiated from hESCs of the three BG lines, human fibroblast feeder HS27 (ATCC), hESC-derived fibroblasts, karyotypically abnormal hESC line BG01 Variant (BG01V) [ ] and EC line NTera2 [ ].

    Techniques: Generated, Expressing

    Diploid pluripotent EC cell line NTera2 and karyotypically abnormal hESC line BG01V can be distinguished from normal hESCs using Illumina array. Comparison of NTera2 and pooled hESC sample resulted a correlation coefficient of 0.8997. Two lists of genes, which were specifically expressed in NTera2 (C) or in hESCs (E) were identified. Likewise, while sharing similarities with BG01 (B, correlation coefficient= 0.9043), BG01V was different from BG01 in expression for many genes, particularly genes from the TGFβ pathway (D, F). Black dots represent genes that were detected at > 0 expression level, blue dots represent genes that were detected both at > 0 expression level and at > 0.99 confidence. Genes plotted outside the two thin red lines were detected at > 2.5- fold difference.

    Journal: BMC Developmental Biology

    Article Title: Genome wide profiling of human embryonic stem cells (hESCs), their derivatives and embryonal carcinoma cells to develop base profiles of U.S. Federal government approved hESC lines

    doi: 10.1186/1471-213X-6-20

    Figure Lengend Snippet: Diploid pluripotent EC cell line NTera2 and karyotypically abnormal hESC line BG01V can be distinguished from normal hESCs using Illumina array. Comparison of NTera2 and pooled hESC sample resulted a correlation coefficient of 0.8997. Two lists of genes, which were specifically expressed in NTera2 (C) or in hESCs (E) were identified. Likewise, while sharing similarities with BG01 (B, correlation coefficient= 0.9043), BG01V was different from BG01 in expression for many genes, particularly genes from the TGFβ pathway (D, F). Black dots represent genes that were detected at > 0 expression level, blue dots represent genes that were detected both at > 0 expression level and at > 0.99 confidence. Genes plotted outside the two thin red lines were detected at > 2.5- fold difference.

    Article Snippet: Samples included 7 hESC lines BG01, BG02, BG03, I6, H1, H7 and H9, EBs that were differentiated from hESCs of the three BG lines, human fibroblast feeder HS27 (ATCC), hESC-derived fibroblasts, karyotypically abnormal hESC line BG01 Variant (BG01V) [ ] and EC line NTera2 [ ].

    Techniques: Expressing

    Differences in the embryoid body (EB) growth rates between the hESC lines I3, I6 and BG01V . hESC colonies were removed from feeder monolayers and grown in low attachment dishes. (A) Phase contrast images of the EBs in the three cell lines cultured in suspension for 10 days. (B) A plot showing differences in the EB growth rate between three cell lines. The growth rate of EBs was calculated by the increase of EB size (total EB area) in each 5 days from day 1 though 25. Values are expressed as percent increase in total areas of EBs (mean ± SEM). Statistical differences for EB growth rates after 10 days in culture between I3 and I6 or BG01V are significant * p

    Journal: BMC Cell Biology

    Article Title: Self-renewal and differentiation capabilities are variable between human embryonic stem cell lines I3, I6 and BG01V

    doi: 10.1186/1471-2121-10-44

    Figure Lengend Snippet: Differences in the embryoid body (EB) growth rates between the hESC lines I3, I6 and BG01V . hESC colonies were removed from feeder monolayers and grown in low attachment dishes. (A) Phase contrast images of the EBs in the three cell lines cultured in suspension for 10 days. (B) A plot showing differences in the EB growth rate between three cell lines. The growth rate of EBs was calculated by the increase of EB size (total EB area) in each 5 days from day 1 though 25. Values are expressed as percent increase in total areas of EBs (mean ± SEM). Statistical differences for EB growth rates after 10 days in culture between I3 and I6 or BG01V are significant * p

    Article Snippet: Cell Culture Human embryonic stem cell lines I3, I6, and BG01V used in this study were cultured on mitomycin C-treated mouse embryonic fibroblasts CF-1 (ATCC, SCRC-1040.2; ).

    Techniques: Cell Culture

    The comparison of the neural progenitor derivation from the I3, I6 and BG01V hESC lines over time under the same neural differentiation-promoting condition . (A) The protocol used to direct hESCs into neural cell lineages. (B-E) Phase contrast images show the progression of the neural induction of hESCs in which the hESC colonies (B) were removed from the MEF feeders and grown into floating aggregates or embryoid bodies (EBs) (C). Neural rosettes (D) were induced by the neural differentiation medium. New cells constantly generated and migrated radially away from the center of the EB and formed a rim of cells (E). Bars in B-E = 100 μm. (F-H) Neural progenitors derived from the I3 cell line. Phase contrast image of I3-differentiated cells (F) were immunostained for nestin (H) and nuclear counterstained with DAPI (G). (I) The plot shows significant differences in the average percent increase in the number of nestin+ cells differentiated from the three cell lines. Values are expressed as percent increase in Nestin+ neural progenitors (mean ± SEM). Statistical analysis shows that the percent increase in number of Nestin+ cells generated from the I3 or I6 cells is significantly greater than that generated from BG01V cells, * p

    Journal: BMC Cell Biology

    Article Title: Self-renewal and differentiation capabilities are variable between human embryonic stem cell lines I3, I6 and BG01V

    doi: 10.1186/1471-2121-10-44

    Figure Lengend Snippet: The comparison of the neural progenitor derivation from the I3, I6 and BG01V hESC lines over time under the same neural differentiation-promoting condition . (A) The protocol used to direct hESCs into neural cell lineages. (B-E) Phase contrast images show the progression of the neural induction of hESCs in which the hESC colonies (B) were removed from the MEF feeders and grown into floating aggregates or embryoid bodies (EBs) (C). Neural rosettes (D) were induced by the neural differentiation medium. New cells constantly generated and migrated radially away from the center of the EB and formed a rim of cells (E). Bars in B-E = 100 μm. (F-H) Neural progenitors derived from the I3 cell line. Phase contrast image of I3-differentiated cells (F) were immunostained for nestin (H) and nuclear counterstained with DAPI (G). (I) The plot shows significant differences in the average percent increase in the number of nestin+ cells differentiated from the three cell lines. Values are expressed as percent increase in Nestin+ neural progenitors (mean ± SEM). Statistical analysis shows that the percent increase in number of Nestin+ cells generated from the I3 or I6 cells is significantly greater than that generated from BG01V cells, * p

    Article Snippet: Cell Culture Human embryonic stem cell lines I3, I6, and BG01V used in this study were cultured on mitomycin C-treated mouse embryonic fibroblasts CF-1 (ATCC, SCRC-1040.2; ).

    Techniques: Generated, Derivative Assay

    Quantitative RT-PCR analysis shows differences in expression levels of neural cell lineage-specific genes between three hES cell lines . The Y-axis plots the fold change for each cell line when compared to its own undifferentiated cells (Day 0). (A) shows a comparison of expression levels of the neural progenitor-specific genes at days 7, 14 and 21 of differentiation. The I3 and I6 cells exhibited higher levels of Nestin and Musashi 1 gene expression than BG01V cells. (B) shows a comparison of the expression levels of the neuronal (MAP2)- and glial (S100B)-specific genes at days 7, 14 and 21 of differentiation. Both of the MAP2 and S100B gene expressions were up-regulated in the I3 and I6 cells.

    Journal: BMC Cell Biology

    Article Title: Self-renewal and differentiation capabilities are variable between human embryonic stem cell lines I3, I6 and BG01V

    doi: 10.1186/1471-2121-10-44

    Figure Lengend Snippet: Quantitative RT-PCR analysis shows differences in expression levels of neural cell lineage-specific genes between three hES cell lines . The Y-axis plots the fold change for each cell line when compared to its own undifferentiated cells (Day 0). (A) shows a comparison of expression levels of the neural progenitor-specific genes at days 7, 14 and 21 of differentiation. The I3 and I6 cells exhibited higher levels of Nestin and Musashi 1 gene expression than BG01V cells. (B) shows a comparison of the expression levels of the neuronal (MAP2)- and glial (S100B)-specific genes at days 7, 14 and 21 of differentiation. Both of the MAP2 and S100B gene expressions were up-regulated in the I3 and I6 cells.

    Article Snippet: Cell Culture Human embryonic stem cell lines I3, I6, and BG01V used in this study were cultured on mitomycin C-treated mouse embryonic fibroblasts CF-1 (ATCC, SCRC-1040.2; ).

    Techniques: Quantitative RT-PCR, Expressing

    Difference in directed neural differentiation between hESC lines I3, I6 and BG01V . Two days after transferring EBs to a poly-D-lysine/laminin substrate, parallel immunocytochemistry and quantitative RT-PCR were performed in hESC-derived cell populations. Immunofluorescent staining for Nestin (A-F) shows that both the I3 and the I6 cells differentiate into the enriched Nestin + neural progenitors while BG01V cells barely generate Nestin + cells. Bars in A-C = 100 μm. (G) qRT-PCR analysis of the gene expression of the neural progenitor markers SOX1 and MSI1, mature neuronal marker MAP2, and astrocyte marker GFAP among the three cell lines at day 17 of differentiation. The Y-axis represents the fold changes of the gene expression for each cell line when compared to its own undifferentiated levels at day 0. The relative levels of these genes expressed by the I3 and I6 cells are higher than those expressed by BG01V cells.

    Journal: BMC Cell Biology

    Article Title: Self-renewal and differentiation capabilities are variable between human embryonic stem cell lines I3, I6 and BG01V

    doi: 10.1186/1471-2121-10-44

    Figure Lengend Snippet: Difference in directed neural differentiation between hESC lines I3, I6 and BG01V . Two days after transferring EBs to a poly-D-lysine/laminin substrate, parallel immunocytochemistry and quantitative RT-PCR were performed in hESC-derived cell populations. Immunofluorescent staining for Nestin (A-F) shows that both the I3 and the I6 cells differentiate into the enriched Nestin + neural progenitors while BG01V cells barely generate Nestin + cells. Bars in A-C = 100 μm. (G) qRT-PCR analysis of the gene expression of the neural progenitor markers SOX1 and MSI1, mature neuronal marker MAP2, and astrocyte marker GFAP among the three cell lines at day 17 of differentiation. The Y-axis represents the fold changes of the gene expression for each cell line when compared to its own undifferentiated levels at day 0. The relative levels of these genes expressed by the I3 and I6 cells are higher than those expressed by BG01V cells.

    Article Snippet: Cell Culture Human embryonic stem cell lines I3, I6, and BG01V used in this study were cultured on mitomycin C-treated mouse embryonic fibroblasts CF-1 (ATCC, SCRC-1040.2; ).

    Techniques: Transferring, Immunocytochemistry, Quantitative RT-PCR, Derivative Assay, Staining, Expressing, Marker

    Digital gene expression of the three undifferentiated hESC lines . It shows that around 92% (10057 out of 10945) of the genes expressed in the I3 cells were also expressed in the BG01V and I6 cells. The large overlap in genes expressed among the 3 hESC lines suggests the presence of a relatively stable core

    Journal: BMC Cell Biology

    Article Title: Self-renewal and differentiation capabilities are variable between human embryonic stem cell lines I3, I6 and BG01V

    doi: 10.1186/1471-2121-10-44

    Figure Lengend Snippet: Digital gene expression of the three undifferentiated hESC lines . It shows that around 92% (10057 out of 10945) of the genes expressed in the I3 cells were also expressed in the BG01V and I6 cells. The large overlap in genes expressed among the 3 hESC lines suggests the presence of a relatively stable core "stemness" transcriptome.

    Article Snippet: Cell Culture Human embryonic stem cell lines I3, I6, and BG01V used in this study were cultured on mitomycin C-treated mouse embryonic fibroblasts CF-1 (ATCC, SCRC-1040.2; ).

    Techniques: Expressing

    Differences in relative expression levels of undifferentiated cell (

    Journal: BMC Cell Biology

    Article Title: Self-renewal and differentiation capabilities are variable between human embryonic stem cell lines I3, I6 and BG01V

    doi: 10.1186/1471-2121-10-44

    Figure Lengend Snippet: Differences in relative expression levels of undifferentiated cell ("stemness") markers among the three undifferentiated hESC lines . (A) Immunofluorescent staining of hESC colonies shows an expression of Oct3/4, NANOG, SSEA4, TRA-1-81 and alkaline phosphatase (AP) activity in all three cell lines. Bar = 100 μm. (B) Quantitative RT-PCR analysis of the relative expression levels of "stemness" genes (NANOG, POU5F1, and UTF1) in three undifferentiated hES cells. The Y-axis plots the fold change of the undifferentiated cell lines in comparison to the undifferentiated I3 cell lines. The expression level of the undifferentiated genes implicates that the I3 hES cells express much less "stemness" (undifferentiated) genes than the I6 and BG01V cell lines. (C) MPSS analysis shows the consistency of the expression levels of the undifferentiated genes (Nanog, POU5F1, and UTF1) compared with the results of the qRT-PCR analysis (B).

    Article Snippet: Cell Culture Human embryonic stem cell lines I3, I6, and BG01V used in this study were cultured on mitomycin C-treated mouse embryonic fibroblasts CF-1 (ATCC, SCRC-1040.2; ).

    Techniques: Expressing, Staining, Activity Assay, Quantitative RT-PCR

    Differences in the proliferation rate between the undifferentiated I3, I6 and BG01V cells . All three cell lines were maintained on the MEF for 4 days post-passaging under the same culture conditions. A four hour BrdU pulse shows significant differences in the proliferation index which is defined as the percentage of BrdU+ nuclei among the total number of propidium iodide (PI)+ cells.(A-I) Images of the three cell line colonies (A, D, G) immunostained for BrdU incorporation (C, F, I) and counterstained with PI (B, E, H). Bars in A, D and G = 100 μm. (J) Bar plot summarizing the differences in diameters of the colonies between the three cell lines. Values are expressed as a percent of the total number of cells (mean ± SEM; BG01V- 554 ± 187 μm, I3 – 184 ± 75 μm, and I6 – 488 ± 165 μm. Sizes of colonies derived from the BG01V or I6 cells are significantly greater than those derived from the I3 cells. ** p

    Journal: BMC Cell Biology

    Article Title: Self-renewal and differentiation capabilities are variable between human embryonic stem cell lines I3, I6 and BG01V

    doi: 10.1186/1471-2121-10-44

    Figure Lengend Snippet: Differences in the proliferation rate between the undifferentiated I3, I6 and BG01V cells . All three cell lines were maintained on the MEF for 4 days post-passaging under the same culture conditions. A four hour BrdU pulse shows significant differences in the proliferation index which is defined as the percentage of BrdU+ nuclei among the total number of propidium iodide (PI)+ cells.(A-I) Images of the three cell line colonies (A, D, G) immunostained for BrdU incorporation (C, F, I) and counterstained with PI (B, E, H). Bars in A, D and G = 100 μm. (J) Bar plot summarizing the differences in diameters of the colonies between the three cell lines. Values are expressed as a percent of the total number of cells (mean ± SEM; BG01V- 554 ± 187 μm, I3 – 184 ± 75 μm, and I6 – 488 ± 165 μm. Sizes of colonies derived from the BG01V or I6 cells are significantly greater than those derived from the I3 cells. ** p

    Article Snippet: Cell Culture Human embryonic stem cell lines I3, I6, and BG01V used in this study were cultured on mitomycin C-treated mouse embryonic fibroblasts CF-1 (ATCC, SCRC-1040.2; ).

    Techniques: Passaging, BrdU Incorporation Assay, Derivative Assay

    Differences in expression of three germ layer markers in the embryoid bodies (EBs) derived from three hES cell lines BG01V, I3 and I6 . (A) Quantitative RT-PCR analysis of expression of keratin C (ectoderm marker) and alpha-Globin (mesoderm marker) at 7 and 14 days in the EBs derived from the three cell lines. The Y-axis plots the fold changes (log scale) in expression levels of the two genes for each cell line when compared to its own undifferentiated levels at Day 0. The I3-derived EBs cultured at days 7 and 14 expressed markedly higher levels of the two genes compared to I6 and BG01V-derived EBs. (B and C) Higher expression of endoderm marker AFP and mesoderm marker IGF-2 gene expression in the I3 than in the BG01V and I6 cell lines revealed by MPSS. Quantitative MPSS analysis of the AFP and IGF-2 in the EBs derived from the hES cell lines BG01V, I3 and I6. The Y-axis plots the fold change (log scale) in expression levels for each cell line when compared to its own undifferentiated cells (Day 0).

    Journal: BMC Cell Biology

    Article Title: Self-renewal and differentiation capabilities are variable between human embryonic stem cell lines I3, I6 and BG01V

    doi: 10.1186/1471-2121-10-44

    Figure Lengend Snippet: Differences in expression of three germ layer markers in the embryoid bodies (EBs) derived from three hES cell lines BG01V, I3 and I6 . (A) Quantitative RT-PCR analysis of expression of keratin C (ectoderm marker) and alpha-Globin (mesoderm marker) at 7 and 14 days in the EBs derived from the three cell lines. The Y-axis plots the fold changes (log scale) in expression levels of the two genes for each cell line when compared to its own undifferentiated levels at Day 0. The I3-derived EBs cultured at days 7 and 14 expressed markedly higher levels of the two genes compared to I6 and BG01V-derived EBs. (B and C) Higher expression of endoderm marker AFP and mesoderm marker IGF-2 gene expression in the I3 than in the BG01V and I6 cell lines revealed by MPSS. Quantitative MPSS analysis of the AFP and IGF-2 in the EBs derived from the hES cell lines BG01V, I3 and I6. The Y-axis plots the fold change (log scale) in expression levels for each cell line when compared to its own undifferentiated cells (Day 0).

    Article Snippet: Cell Culture Human embryonic stem cell lines I3, I6, and BG01V used in this study were cultured on mitomycin C-treated mouse embryonic fibroblasts CF-1 (ATCC, SCRC-1040.2; ).

    Techniques: Expressing, Derivative Assay, Quantitative RT-PCR, Marker, Cell Culture

    (A-C) Morphology of undifferentiated colonies of hESC lines I3, I6, and BG01V . Phase contrast photographs of hESC colonies cultured on mouse embryonic fibroblast feeders for 4 days. The boxes in the center of the colonies indicate areas shown in A1-C1. (A1-C1) High magnification of the center regions of the colonies from each cell line showing distinct morphologies. The colonies of BG01V and I6 lines exhibit highly compact cells with rather vague borders (A1 and C1), while I3 colonies have a mosaic appearance with loosely packed cells (B1). Bars in A and A1 = 100 μm. (D) Three human embryonic stem cell lines display distinct growth characteristics under the same culture conditions. The growth curves show significant differences in percent increase in the number of cells at day 6 in culture between the three cell lines. I3 cells grow slower than the other two cell lines and have a tendency to differentiate. Therefore, it is more difficult to maintain the I3 cell line in an undifferentiated state. In contrast, the I6 and BG01V cell lines grow faster and are passaged 2–3 days earlier than the I3 cells. Statistical differences for percent increases in cell numbers at 6 days between the BG01V or I6 and I3 are significant ** p

    Journal: BMC Cell Biology

    Article Title: Self-renewal and differentiation capabilities are variable between human embryonic stem cell lines I3, I6 and BG01V

    doi: 10.1186/1471-2121-10-44

    Figure Lengend Snippet: (A-C) Morphology of undifferentiated colonies of hESC lines I3, I6, and BG01V . Phase contrast photographs of hESC colonies cultured on mouse embryonic fibroblast feeders for 4 days. The boxes in the center of the colonies indicate areas shown in A1-C1. (A1-C1) High magnification of the center regions of the colonies from each cell line showing distinct morphologies. The colonies of BG01V and I6 lines exhibit highly compact cells with rather vague borders (A1 and C1), while I3 colonies have a mosaic appearance with loosely packed cells (B1). Bars in A and A1 = 100 μm. (D) Three human embryonic stem cell lines display distinct growth characteristics under the same culture conditions. The growth curves show significant differences in percent increase in the number of cells at day 6 in culture between the three cell lines. I3 cells grow slower than the other two cell lines and have a tendency to differentiate. Therefore, it is more difficult to maintain the I3 cell line in an undifferentiated state. In contrast, the I6 and BG01V cell lines grow faster and are passaged 2–3 days earlier than the I3 cells. Statistical differences for percent increases in cell numbers at 6 days between the BG01V or I6 and I3 are significant ** p

    Article Snippet: Cell Culture Human embryonic stem cell lines I3, I6, and BG01V used in this study were cultured on mitomycin C-treated mouse embryonic fibroblasts CF-1 (ATCC, SCRC-1040.2; ).

    Techniques: Cell Culture

    BG01V embryonic stem cells differentiate into neural progenitors (A) Contrast phase microscopic image of ESC grown on a MEF feeder layer (left), and generated NP grown on laminin (right). Arrows point to ESC colonies. (B) Cell lysates were prepared, and the expression of the indicated markers was analyzed by Western blotting as described in the M M section. β-tubulin was used as a loading control. (C) Immunocytochemical staining of stem cells and differentiated neural progenitors with antibodies (Ab) against the cell specific markers (red/green) as indicated, and DAPI staining (blue) was performed as described in the M M section. Differentiation experiments were repeated six times with the consistent results.

    Journal: Stem cells (Dayton, Ohio)

    Article Title: Nuclear Factor I isoforms regulate gene expression during the differentiation of human neural progenitors to astrocytes

    doi: 10.1002/stem.35

    Figure Lengend Snippet: BG01V embryonic stem cells differentiate into neural progenitors (A) Contrast phase microscopic image of ESC grown on a MEF feeder layer (left), and generated NP grown on laminin (right). Arrows point to ESC colonies. (B) Cell lysates were prepared, and the expression of the indicated markers was analyzed by Western blotting as described in the M M section. β-tubulin was used as a loading control. (C) Immunocytochemical staining of stem cells and differentiated neural progenitors with antibodies (Ab) against the cell specific markers (red/green) as indicated, and DAPI staining (blue) was performed as described in the M M section. Differentiation experiments were repeated six times with the consistent results.

    Article Snippet: Human BG01V embryonic stem cells (ATCC, Rockville, MD) were cultured on mitomycin C-inactivated mouse embryonic fibroblasts (MEF) layer.

    Techniques: Generated, Expressing, Western Blot, Staining