Differentiation and Molecular Properties of Mesenchymal Stem Cells Derived from Murine Induced Pluripotent Stem Cells Derived on Gelatin or Collagen

Figure 1

Derivation of mouse iPSC-derived mesenchymal stem cells (iMSC). (a) Protocol for derivation of iMSCs. iPSCs were maintained on mouse embryonic fibroblast (MEF) in iPS medium with leukemia inhibitory factor (LIF) and harvested and dissociated into single cells that were seeded into gelatin ①- or collagen ②-coated plates. After 1 day, the iPS medium was supplemented with an equal volume of the derivation medium with dexamethasone and ascorbic acid, and after 2 days, the medium was replaced with derivation medium. After 8 days, cells (passage zero; P0) were harvested and replaced in new gelatin- or collagen-coated plates in expansion medium. (b) Adherent spindle-shaped fibroblast-like cells at P0 appeared on 8-derivation day. (c) Homogenous fibroblast-like cells at P3. (d) Characterization of iMSCs on gelatin (iMSCs/G) and on collagen (iMSCs/C) at P4 to P7. Bone marrow (BM) MSC at P5 was provided by cell sorting from mouse bone marrow. The gray histogram represents the negative control staining with fluorescence-conjugated isotype IgG and white overlay represents antigen at CD90.2, CD105, CD73, CD29, CD44, CD45, Sca-1, and CD140α.