Retinal Lineage Therapeutic Specific Effect of Human Orbital and Abdominal Adipose-Derived Mesenchymal Stem Cells

Figure 3

Differentiation of OASCs to RPE like cells. (a, b) Coculture of p-RPE cells and OASCs. (a) Prestaining of cells: OASCs were marked with CSFE (green color), and RPE mature cells were marked with Violet Cell Trace (blue color). (b) FACS sorter: three distinct populations are defined after coculture: RPE cells, OASCs and OASCs, and RPE cells which were fused together. Autofluorescent cells were excluded according to negative and positive controls of labeled cells. (c) Evidence of differentiation of OASCs to RPE like cells by RT-qPCR. Results are presented as relative to sample plot. After coculture with RPE cells, OASCs exhibited an increase in transcription of early and late RPE markers. (d) Evidence of differentiation potential of OASCs by immunofluorescence. (A) Sorted OASCs derived RPE-like cells labeled with CFSE showing nuclear localized OTX2 expression. (B) Sorted OASCs derived RPE-like cells showing nuclear localized PAX6 expression (C) Positive control of sorted RPE cells in Violet Cell Trace showing nuclear localized OTX2 expression. (D) Positive control of sorted RPE cells in Violet Cell Trace showing nuclear localized PAX6 expression. (E, F) Negative control of OASCs that did not show any OTX2 expression or PAX6 expression. CT: cell trace. Scale bar: 100 μm. .

(a)

(b)

(c)

(d)