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. 2015 Feb 26;4(3):459–472. doi: 10.1016/j.stemcr.2015.01.017

Figure 5.

Figure 5

WNT3A and FGF2 Maintain MSC Characteristics in Multiple Passages

(A) Cell morphology of MSCs expanded as indicated. The scale bar represents 200 μm.

(B) Flow cytometry analysis of WF-MSCs (+WNT+FGF) and F-MSCs (+FGF). The percentage represents the number of positive cells for the indicated surface marker. Values are the mean of two MSC donors. NC, negative control.

(C) Schematic representation of the working plan for the multilineage differentiation.

(D) Von Kossa staining (calcium phosphate deposits) and ALP mRNA levels of MSCs expanded in the indicated conditions and subsequently differentiated in osteogenic medium. The scale bar represents 150 μm.

(E) Oil red O staining (lipid accumulation) and FABP4 mRNA levels of MSCs expanded in the indicated conditions and subsequently differentiated in adipogenic medium. The scale bar represents 100 μm.

Transcript analysis in (D) and (E) was performed on P4 expanded cells. Images of P1 cells are representative of two independent MSC donors; P4 images are representative of three MSC donors. ∗∗p < 0.01.

(F) Gene expression profile of MSCs expanded in the indicated conditions prior to differentiation. Values represent means ± SEM. The data are representative of three MSC donors with two biological replicates per donor. ∗p < 0.05, ∗∗p < 0.01.