Figure 2.

Induced pluripotent stem cells (iPSCs) derived from gene-targeted mesenchymal stem cells (MSCs). (a) Southern blot identification of iPSC clones targeted at COL1A2. EV, EcoR V; neo, neomycin resistance gene; pA, polyadenylation site; S, stop codons; L, loxP site; OI12, parental OIMSC12 culture; Poly, polyclonal G418-resistant MSCs transduced with AAV vector. (b) Identification of targeted allele by complementary DNA (cDNA) sequencing in two gene-targeted clones, with controls of untransduced OI12 iPSCs (iPSC-OI12-FVc3) and a random integrant. TW, wild-type allele targeted; TM, mutant allele targeted. (c) Pluripotency gene expression as measured by microarray. (d) Teratoma assay of iPSCe2-FVc3. H&E, hematoxylin and eosin; DAPI, 4,6-diamidino-2-phenylindole; MAP-2, microtubule associated protein-2; AFP, α-fetoprotein; SMA, α-smooth muscle actin. Bar = 50 µm.