Epithelial-Mesenchymal Transition Promotes the Differentiation Potential of Xenopus tropicalis Immature Sertoli Cells

Figure 5

In vitro differentiation of XtiSCs. XtiSCs were treated with FGF2, CHIR99021, or 0.1% DMSO as a control for 3-4 days followed by the replacement of medium with osteogenic and chondrogenic induction media. (a) Osteocyte differentiation of XtiSCs was evidenced as a calcium deposit reacting with alizarin red (A). Cells were lysed at low pH to release alizarin red dye from the stained monolayer for a colorimetric test as illustrated in diagram (B). (b) Alcian blue was used to stain glycosaminoglycans in cartilages. In the control (untreated cells), the dye reacts only with the periphery of cell clusters. On the other hand, alcian blue staining is strong and uniform in XtiSC clumps after FGF2 and CHIR99021 treatments, indicating the high level of proteoglycan forming the extracellular matrix. However, collagen type II, a specific marker of the cartilage matrix, was expressed in cells pretreated with CHIR99021 only. DAPI was used as cell counterstains. Results are representative of three biological replicates. Scale bar: 100 μm for bright field images and 50 μm for fluorescent images (lower panel of (b)); , .

(a)

(b)