Characterization of Mesenchymal Stem Cell-Like Cells Derived From Human iPSCs via Neural Crest Development and Their Application for Osteochondral Repair

Figure 6

In vitro development of tissue-engineered tissue from iNCMSCs. (a–c) The gross appearance of iNCMSC-TEC developed in 35 mm dishes at day 7. Thin monolayer cell sheet was formed in the culture bottom surface after 7 days of high-density culture (a), and once artificially detached (black arrows), a thick sheet-like construct (iNCMSC-TEC) developed through active tissue contraction within a few minutes (b). HE staining and immunostaining for COL1 and COL2 of transverse section are shown in their corresponding right panels. Scale bars = 50 μm. The iNCMSC-TEC exhibited sufficient strength to handle with forceps (c). (d) Ex vivo implantation of iNCMSC-TEC into osteochondral defects of cadaver rats. iNCMSC-TEC readily filled the defect created in the femoral groove (white arrows). Scale bars = 1 mm. (e, f) Chondrogenic potential of iNCMSC-TEC. COL2 immunostaining (e) and Safranin-O staining (f) of iNCMSC-TEC cultured in chondrogenic medium for 1 month. Scale bars = 500 μm. Abbreviations: HE: hematoxylin and eosin staining; COL1: type 1 collagen; COL2: type 2 collagen; Saf-O: Safranin-O.

(a)

(b)

(c)

(d)

(e)

(f)