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. 2015 Sep 24;5(4):647–659. doi: 10.1016/j.stemcr.2015.08.015

Figure 5.

Figure 5

Validation of Neural Differentiation and Gene Targeting and Preparation of the Human iPSC MCB and WCBs

(A and B) Use of iPSCs (LiPSC-TR1.2) generated using a cGMP-compatible process in pre-clinical studies for neural differentiation (A) and genetic engineering (B).

(A, a–c) Neural rosettes formed via EBs were isolated manually and expanded to a homogenous NSC population. Immunocytochemical analysis showed positive expression of the NSC markers SOX1 and NESTIN. Scale bars, 200 μm, except in b and c (100 μm).

(B, a–c) TALEN-mediated homologous recombination targeting the safe harbor site AAVS1 on chromosome 19. A representative example of a GFP-positive clone is shown. GFP was driven by the constitutively active CAG promoter. Scale bars, 200 μm.

(C) Human iPSCs generated under cGMP conditions can be used as MCB seed stocks to create working cell banks under both the GMP setting (for manufacturing specialized cell therapy products) and in a non-GMP environment (to carry out research studies for multiple cell therapy applications).