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Fig. 3 | Stem Cell Research & Therapy

Fig. 3

From: In vitro generation of Sertoli-like and haploid spermatid-like cells from human umbilical cord perivascular cells

Fig. 3

First trimester (FTM) and term human umbilical cord perivascular cells (HUCPVCs) differentiate towards both Sertoli- and germ-like cells within the same culture environment. a Undifferentiated FTM HUCPVCs and FTM HUCPVCs at the end of Step 1, Step 2, and Step 3. b FTM HUCPVCs (left) and term HUCPVCs (right) at Step 3 of the differentiation protocol showing adherent cells (black arrow) and small round loosely attached cells (white arrow). c RT-PCR for Sertoli-associated gene expression including follicle stimulating hormone receptor (FSHR), anti-müllerian hormone (AMH), SOX9, clusterin (CLU), and transforming growth factor receptor beta (TGFRβ1) in FTM HUCPVCs and term HUCPVCs at the undifferentiated stage (UD), Step 1 (S1), Step 2 (S2), and Step 3 (S3); granulosa cells used as positive control (+). – indicates no cDNA controls. d Undifferentiated (UNDIFF) FTM HUCPVCs (negative control) (top, left) and term HUCPVCs (bottom, left) (negative control) immunostained for FSHR (green, undetected) and counterstained with Hoechst to show live nuclei. Granulosa cells (GLCs) were used as a positive control. e Flow cytometric quantification of the percentage of FTM HUCPVCs that upregulated FSHR at the end of Step 2 when compared to undifferentiated controls. f RT-PCR for early germ cell-associated gene expression of VASA and DAZL in the same samples as panel c. g RT-PCR for GAPDH results shown in panels c and f. h Undifferentiated FTM HUCPVCs (left) immunostained for DAZL (green, undetected) and counterstained with Hoechst to show live nuclei. i Undifferentiated FTM HUCPVCs (top, left) and term HUCPVCs (bottom, left) immunostained for VASA (green, undetected) and counterstained with Hoechst. j Flow cytometric quantification of the percentage of FTM HUCPVCs that upregulated VASA at the end of Step 2 when compared to undifferentiated controls

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