Cell Culture:
Modification:
other:
Expressing:Article Title: Characterization, Preconditioning, Safety, and Other Issues of MSC-Derived EVs and Secretome
Article Snippet: AT-MSCs [ ] , hTERT-immortalized AT-MSCs (SCRC-4000TM cell line, purchased from ATCC ® ), adapted to suspension culture (S-hMSCs); S-hMSCs had comparable growth kinetics to primary and immortalized AT-MSCs; doubling time of app. 55 h; retained close to 90% of CD73 and CD105 expression levels, the CD90 receptor being downregulated during the adherent to suspension adaptation process, upregulation of the transcripts coding for CD44, CD46, and CD47 compared to the expression levels in primary and immortalized AT-MSCs (by RNA sequencing) , EVs isolated via ultracentrifugation; mean size 153 nm; expressing CD63, CD81, TSG101; functional studies are planned , Not yet performed. .. AT-MSCs [ ] , iMSCs purchased from ATCC ® (SCRC-4000 TM ); showed positive expression of CD29, CD44, CD73, CD90, CD105, absence of CD31, CD34, CD45, and HLA-DR; differentiation into adipocytes, osteocytes, and chondrocytes confirmed , Isolated using the Total Exosome Isolation Reagent (4478359, Invitrogen, Waltham, MA, USA) from iMSCs primed with serum derived from a non-human primate rheumatoid arthritis (RA) model; serum IL-4 and the proportion of Th2 (CD4+CD25+GATA3+) and M2 (CD11c-CD206+ of CD45+CD64+) cells were significantly increased compared to the control when sEVs-RA were administered to the collagen-induced arthritis model; sEVs-RA alleviated cartilage damage by significantly lowering the concentrations of pro-inflammatory cytokines (TNF-α, keratinocyte chemoattractant, IL-12p70) , Was not performed. .. AT-MSCs [ , ] , Expression of hTERT mRNA; nuclear hTERT immunoreaction; more than PD100, normal karyotype (with a clone with chromosomal aberrations after PD83), no signs of senescence by SA-bGal staining; retained expression of CD44, CD73, CD90, CD105, did not express CD3, CD14, CD19, CD34, CD45, HLA-DR; osteogenic and adipogenic differentiation; in vivo tumorigenicity was absent , Exo-spin Kit (EX01, Cell Guidance Sytems Ltd., Cambridge, UK) or ultracentrifugation followed by the PEG precipitation method applied for the enrichment of sEVs; remarkable number of sEVs with proper size (40–150 nm) and protein concentration produced, proper integrity, size distribution, and polydispersity index; positive for common exosomal markers, including CD63 and TSG101 , Was not performed.
Isolation:Article Title: Characterization, Preconditioning, Safety, and Other Issues of MSC-Derived EVs and Secretome
Article Snippet: AT-MSCs [ ] , hTERT-immortalized AT-MSCs (SCRC-4000TM cell line, purchased from ATCC ® ), adapted to suspension culture (S-hMSCs); S-hMSCs had comparable growth kinetics to primary and immortalized AT-MSCs; doubling time of app. 55 h; retained close to 90% of CD73 and CD105 expression levels, the CD90 receptor being downregulated during the adherent to suspension adaptation process, upregulation of the transcripts coding for CD44, CD46, and CD47 compared to the expression levels in primary and immortalized AT-MSCs (by RNA sequencing) , EVs isolated via ultracentrifugation; mean size 153 nm; expressing CD63, CD81, TSG101; functional studies are planned , Not yet performed. .. AT-MSCs [ ] , iMSCs purchased from ATCC ® (SCRC-4000 TM ); showed positive expression of CD29, CD44, CD73, CD90, CD105, absence of CD31, CD34, CD45, and HLA-DR; differentiation into adipocytes, osteocytes, and chondrocytes confirmed , Isolated using the Total Exosome Isolation Reagent (4478359, Invitrogen, Waltham, MA, USA) from iMSCs primed with serum derived from a non-human primate rheumatoid arthritis (RA) model; serum IL-4 and the proportion of Th2 (CD4+CD25+GATA3+) and M2 (CD11c-CD206+ of CD45+CD64+) cells were significantly increased compared to the control when sEVs-RA were administered to the collagen-induced arthritis model; sEVs-RA alleviated cartilage damage by significantly lowering the concentrations of pro-inflammatory cytokines (TNF-α, keratinocyte chemoattractant, IL-12p70) , Was not performed. .. AT-MSCs [ , ] , Expression of hTERT mRNA; nuclear hTERT immunoreaction; more than PD100, normal karyotype (with a clone with chromosomal aberrations after PD83), no signs of senescence by SA-bGal staining; retained expression of CD44, CD73, CD90, CD105, did not express CD3, CD14, CD19, CD34, CD45, HLA-DR; osteogenic and adipogenic differentiation; in vivo tumorigenicity was absent , Exo-spin Kit (EX01, Cell Guidance Sytems Ltd., Cambridge, UK) or ultracentrifugation followed by the PEG precipitation method applied for the enrichment of sEVs; remarkable number of sEVs with proper size (40–150 nm) and protein concentration produced, proper integrity, size distribution, and polydispersity index; positive for common exosomal markers, including CD63 and TSG101 , Was not performed.
Derivative Assay:Article Title: Characterization, Preconditioning, Safety, and Other Issues of MSC-Derived EVs and Secretome
Article Snippet: AT-MSCs [ ] , hTERT-immortalized AT-MSCs (SCRC-4000TM cell line, purchased from ATCC ® ), adapted to suspension culture (S-hMSCs); S-hMSCs had comparable growth kinetics to primary and immortalized AT-MSCs; doubling time of app. 55 h; retained close to 90% of CD73 and CD105 expression levels, the CD90 receptor being downregulated during the adherent to suspension adaptation process, upregulation of the transcripts coding for CD44, CD46, and CD47 compared to the expression levels in primary and immortalized AT-MSCs (by RNA sequencing) , EVs isolated via ultracentrifugation; mean size 153 nm; expressing CD63, CD81, TSG101; functional studies are planned , Not yet performed. .. AT-MSCs [ ] , iMSCs purchased from ATCC ® (SCRC-4000 TM ); showed positive expression of CD29, CD44, CD73, CD90, CD105, absence of CD31, CD34, CD45, and HLA-DR; differentiation into adipocytes, osteocytes, and chondrocytes confirmed , Isolated using the Total Exosome Isolation Reagent (4478359, Invitrogen, Waltham, MA, USA) from iMSCs primed with serum derived from a non-human primate rheumatoid arthritis (RA) model; serum IL-4 and the proportion of Th2 (CD4+CD25+GATA3+) and M2 (CD11c-CD206+ of CD45+CD64+) cells were significantly increased compared to the control when sEVs-RA were administered to the collagen-induced arthritis model; sEVs-RA alleviated cartilage damage by significantly lowering the concentrations of pro-inflammatory cytokines (TNF-α, keratinocyte chemoattractant, IL-12p70) , Was not performed. .. AT-MSCs [ , ] , Expression of hTERT mRNA; nuclear hTERT immunoreaction; more than PD100, normal karyotype (with a clone with chromosomal aberrations after PD83), no signs of senescence by SA-bGal staining; retained expression of CD44, CD73, CD90, CD105, did not express CD3, CD14, CD19, CD34, CD45, HLA-DR; osteogenic and adipogenic differentiation; in vivo tumorigenicity was absent , Exo-spin Kit (EX01, Cell Guidance Sytems Ltd., Cambridge, UK) or ultracentrifugation followed by the PEG precipitation method applied for the enrichment of sEVs; remarkable number of sEVs with proper size (40–150 nm) and protein concentration produced, proper integrity, size distribution, and polydispersity index; positive for common exosomal markers, including CD63 and TSG101 , Was not performed.
Control:Article Title: Characterization, Preconditioning, Safety, and Other Issues of MSC-Derived EVs and Secretome
Article Snippet: AT-MSCs [ ] , hTERT-immortalized AT-MSCs (SCRC-4000TM cell line, purchased from ATCC ® ), adapted to suspension culture (S-hMSCs); S-hMSCs had comparable growth kinetics to primary and immortalized AT-MSCs; doubling time of app. 55 h; retained close to 90% of CD73 and CD105 expression levels, the CD90 receptor being downregulated during the adherent to suspension adaptation process, upregulation of the transcripts coding for CD44, CD46, and CD47 compared to the expression levels in primary and immortalized AT-MSCs (by RNA sequencing) , EVs isolated via ultracentrifugation; mean size 153 nm; expressing CD63, CD81, TSG101; functional studies are planned , Not yet performed. .. AT-MSCs [ ] , iMSCs purchased from ATCC ® (SCRC-4000 TM ); showed positive expression of CD29, CD44, CD73, CD90, CD105, absence of CD31, CD34, CD45, and HLA-DR; differentiation into adipocytes, osteocytes, and chondrocytes confirmed , Isolated using the Total Exosome Isolation Reagent (4478359, Invitrogen, Waltham, MA, USA) from iMSCs primed with serum derived from a non-human primate rheumatoid arthritis (RA) model; serum IL-4 and the proportion of Th2 (CD4+CD25+GATA3+) and M2 (CD11c-CD206+ of CD45+CD64+) cells were significantly increased compared to the control when sEVs-RA were administered to the collagen-induced arthritis model; sEVs-RA alleviated cartilage damage by significantly lowering the concentrations of pro-inflammatory cytokines (TNF-α, keratinocyte chemoattractant, IL-12p70) , Was not performed. .. AT-MSCs [ , ] , Expression of hTERT mRNA; nuclear hTERT immunoreaction; more than PD100, normal karyotype (with a clone with chromosomal aberrations after PD83), no signs of senescence by SA-bGal staining; retained expression of CD44, CD73, CD90, CD105, did not express CD3, CD14, CD19, CD34, CD45, HLA-DR; osteogenic and adipogenic differentiation; in vivo tumorigenicity was absent , Exo-spin Kit (EX01, Cell Guidance Sytems Ltd., Cambridge, UK) or ultracentrifugation followed by the PEG precipitation method applied for the enrichment of sEVs; remarkable number of sEVs with proper size (40–150 nm) and protein concentration produced, proper integrity, size distribution, and polydispersity index; positive for common exosomal markers, including CD63 and TSG101 , Was not performed.
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