Reprogrammed Cells Display Distinct Proteomic Signatures Associated with Colony Morphology Variability

Figure 2

Colony morphology is associated with the capacity to form EB and directed differentiation. (a) An overview of the EB formation protocol. (b) Phase-contrast images of the spontaneously differentiated EB at the following time points; Day 2 and Day 10 in suspension plates and Day 2 and Day 14 on Matrigel-coated plates. (c) A score card of the ability to form the different embryonic germ layers for 14 selected cell lines. The analysis was performed at the end of the protocol (Day 29) and analyzed by immunohistochemistry for the markers TUJ1 (ectoderm), AFP (endoderm), and SMA (mesoderm). (d) An overview of the directed differentiation protocol towards definite endoderm (DE) and primitive gut tube (PG). (e) The directed differentiation capacity as measured with flow cytometry analysis for the pluripotency marker POU5F1 and the DE marker SOX17 in line 7-B (stable colony), line 4-C (unstable class 1), and line 4-A (unstable class 2). There were no SOX17+ cells in line 4-A at any of the time points.

(a) Spontaneous EB differentiation

(b)

(c) EB score card

(d) Directed differentiation

(e)