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. 2024 Nov 14;10:468. doi: 10.1038/s41420-024-02231-9

Fig. 1. The optimization and characterization of granulosa-like cell differentiation protocols from human induced pluripotent stem cells (iPSCs).

Fig. 1

A Schematic representing of a three-stage approach (mesoderm, intermediate mesoderm (IM), granulosa cells (GCs)) to generate patient-specific models of GC-like cells by guided differentiation of iPSCs. B mRNA expression levels of key genes in stem cell pluripotency (differentiation day 0), mesoderm (differentiation day 2), and intermediate mesoderm cells (differentiation day 4–6) by using qRT-PCR analysis under different combinations of SB4 compound and/or BMP4. C A representative table depicting small molecular chemicals and recombinant proteins used in four tested conditions of GC differentiation in this study. D Brightfield representative images showing the intermediate mesoderm embryonic bodies (EBs) before attachments (day 6) and as differentiated GC (day 12). E Comparative examination of mRNA expression levels of mesoderm, IM, GC marker genes between four different differentiation conditions. F Immunofluorescence (IF) staining characterizations of mesoderm marker (GATA4) and GC markers (AMHR2, CYP19A1) in differentiation condition #2. The luteinized cumulus granulosa cells served as IF positive control cells. Scale bar, 50 μm. G The quantification of fluorescence images of (F) (left panel). Bars indicate the mean ± SD (n = 4).