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bPGCLC induction. (A) Scheme for the direct induction of bPGCLCs from bES cells. (B) PGCLC induction with a WNT agonist plus WNT antagonist. Shown are bright-field (BF), fluorescence images and FACS analysis of D4 aggregates of bES cells. BT, BLIMP1-tdTomato. TN, <t>TFAP2C-mNeonGreen.</t> Scale bar: 200 µm. (C) Scheme for the induction of bPGCLCs under the culture conditions indicated. (D) bES cells after 24 h of incubation under the conditions indicated. (E) PGCLC induction with BMP4 and a WNT antagonist. Shown are bright-field (BF) and fluorescence images and FACS patterns of D4 aggregates of bES cells after 24 h of pre-incubation. BT, BLIMP1-tdTomato. TN, TFAP2C-mNeonGreen. Scale bar: 200 µm. (F) PGCLC induction from various cell lines. C1-144 yielded the most BTTN-positive cells when the BMP4 concentration was 200 ng/ml, while C11-928 and N5-319 were best when the BMP4 concentration was 50 ng/ml. The concentration of CHIR was 6 µM in the all experiments. FACS analyses were done on day 4 of PGCLC induction. BT, BLIMP1-tdTomato. TN, TFAP2C-mNeonGreen. Scale bar: 200 µm.
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bPGCLC induction. (A) Scheme for the direct induction of bPGCLCs from bES cells. (B) PGCLC induction with a WNT agonist plus WNT antagonist. Shown are bright-field (BF), fluorescence images and FACS analysis of D4 aggregates of bES cells. BT, BLIMP1-tdTomato. TN, <t>TFAP2C-mNeonGreen.</t> Scale bar: 200 µm. (C) Scheme for the induction of bPGCLCs under the culture conditions indicated. (D) bES cells after 24 h of incubation under the conditions indicated. (E) PGCLC induction with BMP4 and a WNT antagonist. Shown are bright-field (BF) and fluorescence images and FACS patterns of D4 aggregates of bES cells after 24 h of pre-incubation. BT, BLIMP1-tdTomato. TN, TFAP2C-mNeonGreen. Scale bar: 200 µm. (F) PGCLC induction from various cell lines. C1-144 yielded the most BTTN-positive cells when the BMP4 concentration was 200 ng/ml, while C11-928 and N5-319 were best when the BMP4 concentration was 50 ng/ml. The concentration of CHIR was 6 µM in the all experiments. FACS analyses were done on day 4 of PGCLC induction. BT, BLIMP1-tdTomato. TN, TFAP2C-mNeonGreen. Scale bar: 200 µm.
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bPGCLC induction. (A) Scheme for the direct induction of bPGCLCs from bES cells. (B) PGCLC induction with a WNT agonist plus WNT antagonist. Shown are bright-field (BF), fluorescence images and FACS analysis of D4 aggregates of bES cells. BT, BLIMP1-tdTomato. TN, <t>TFAP2C-mNeonGreen.</t> Scale bar: 200 µm. (C) Scheme for the induction of bPGCLCs under the culture conditions indicated. (D) bES cells after 24 h of incubation under the conditions indicated. (E) PGCLC induction with BMP4 and a WNT antagonist. Shown are bright-field (BF) and fluorescence images and FACS patterns of D4 aggregates of bES cells after 24 h of pre-incubation. BT, BLIMP1-tdTomato. TN, TFAP2C-mNeonGreen. Scale bar: 200 µm. (F) PGCLC induction from various cell lines. C1-144 yielded the most BTTN-positive cells when the BMP4 concentration was 200 ng/ml, while C11-928 and N5-319 were best when the BMP4 concentration was 50 ng/ml. The concentration of CHIR was 6 µM in the all experiments. FACS analyses were done on day 4 of PGCLC induction. BT, BLIMP1-tdTomato. TN, TFAP2C-mNeonGreen. Scale bar: 200 µm.
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Image Search Results


bPGCLC induction. (A) Scheme for the direct induction of bPGCLCs from bES cells. (B) PGCLC induction with a WNT agonist plus WNT antagonist. Shown are bright-field (BF), fluorescence images and FACS analysis of D4 aggregates of bES cells. BT, BLIMP1-tdTomato. TN, TFAP2C-mNeonGreen. Scale bar: 200 µm. (C) Scheme for the induction of bPGCLCs under the culture conditions indicated. (D) bES cells after 24 h of incubation under the conditions indicated. (E) PGCLC induction with BMP4 and a WNT antagonist. Shown are bright-field (BF) and fluorescence images and FACS patterns of D4 aggregates of bES cells after 24 h of pre-incubation. BT, BLIMP1-tdTomato. TN, TFAP2C-mNeonGreen. Scale bar: 200 µm. (F) PGCLC induction from various cell lines. C1-144 yielded the most BTTN-positive cells when the BMP4 concentration was 200 ng/ml, while C11-928 and N5-319 were best when the BMP4 concentration was 50 ng/ml. The concentration of CHIR was 6 µM in the all experiments. FACS analyses were done on day 4 of PGCLC induction. BT, BLIMP1-tdTomato. TN, TFAP2C-mNeonGreen. Scale bar: 200 µm.

Journal: The Journal of Reproduction and Development

Article Title: Efficient derivation of embryonic stem cells and primordial germ cell-like cells in cattle

doi: 10.1262/jrd.2023-087

Figure Lengend Snippet: bPGCLC induction. (A) Scheme for the direct induction of bPGCLCs from bES cells. (B) PGCLC induction with a WNT agonist plus WNT antagonist. Shown are bright-field (BF), fluorescence images and FACS analysis of D4 aggregates of bES cells. BT, BLIMP1-tdTomato. TN, TFAP2C-mNeonGreen. Scale bar: 200 µm. (C) Scheme for the induction of bPGCLCs under the culture conditions indicated. (D) bES cells after 24 h of incubation under the conditions indicated. (E) PGCLC induction with BMP4 and a WNT antagonist. Shown are bright-field (BF) and fluorescence images and FACS patterns of D4 aggregates of bES cells after 24 h of pre-incubation. BT, BLIMP1-tdTomato. TN, TFAP2C-mNeonGreen. Scale bar: 200 µm. (F) PGCLC induction from various cell lines. C1-144 yielded the most BTTN-positive cells when the BMP4 concentration was 200 ng/ml, while C11-928 and N5-319 were best when the BMP4 concentration was 50 ng/ml. The concentration of CHIR was 6 µM in the all experiments. FACS analyses were done on day 4 of PGCLC induction. BT, BLIMP1-tdTomato. TN, TFAP2C-mNeonGreen. Scale bar: 200 µm.

Article Snippet: Existing targeting vectors were treated with restriction enzymes, and mNeonGreen sequences were amplified by PCR from LifeAct-mNeonGreen (98877; Addgene) and cloned using an in-fusion HD cloning kit (TaKaRa Bio, Shiga, Japan).

Techniques: Fluorescence, Incubation, Concentration Assay

Separation of bPGCLCs by surface markers. (A) FACS analysis of surface marker protein expression in BTTN-positive and -negative cells in D6 aggregate. Red and grey histograms show the cells with antibody and without antibody, respectively. BT, BLIMP1-tdTomato. TN, TFAP2C-mNeonGreen. (B) Separation of induced PGCLCs (C1-144 ES cell) by surface markers KIT/CD117 and CD44. BT, BLIMP1-tdTomato. TN, TFAP2C-mNeonGreen. (C) Separation of PGCLCs derived from non-reporter bES cells by surface markers. (D) Comparison of gene expression in BTTN+ cells and KIT+/CD44– cells. Shown are ΔC t values determined by qPCR analysis.

Journal: The Journal of Reproduction and Development

Article Title: Efficient derivation of embryonic stem cells and primordial germ cell-like cells in cattle

doi: 10.1262/jrd.2023-087

Figure Lengend Snippet: Separation of bPGCLCs by surface markers. (A) FACS analysis of surface marker protein expression in BTTN-positive and -negative cells in D6 aggregate. Red and grey histograms show the cells with antibody and without antibody, respectively. BT, BLIMP1-tdTomato. TN, TFAP2C-mNeonGreen. (B) Separation of induced PGCLCs (C1-144 ES cell) by surface markers KIT/CD117 and CD44. BT, BLIMP1-tdTomato. TN, TFAP2C-mNeonGreen. (C) Separation of PGCLCs derived from non-reporter bES cells by surface markers. (D) Comparison of gene expression in BTTN+ cells and KIT+/CD44– cells. Shown are ΔC t values determined by qPCR analysis.

Article Snippet: Existing targeting vectors were treated with restriction enzymes, and mNeonGreen sequences were amplified by PCR from LifeAct-mNeonGreen (98877; Addgene) and cloned using an in-fusion HD cloning kit (TaKaRa Bio, Shiga, Japan).

Techniques: Marker, Expressing, Derivative Assay, Comparison, Gene Expression