Cord blood cell-derived iPSCs as a new candidate for chondrogenic differentiation and cartilage regeneration

Abstract

BackgroundThe native articular cartilage lacks the ability to heal. Currently, ex vivo expanded chondrocytes or bone marrow-derived mesenchymal stem cells are used to regenerate the damaged cartilage. With unlimited self-renewal ability and multipotency, human induced pluripotent stem cells (hiPSCs) have been highlighted as a new replacement cell source for cartilage repair. Still, further research is needed on cartilage regeneration using cord blood mononuclear cell-derived hiPSCs (CBMC-hiPSCs).MethodsHuman iPSCs were generated from CBMCs using the Sendai virus. The characterization of CBMC-hiPSCs was performed by various assays. Embryonic bodies (EBs) were obtained using CBMC-hiPSCs, and outgrowth cells were induced by plating the EBs onto a gelatin-coated plate. Expanded outgrowth cells were detached and dissociated for chondrogenic differentiation. Outgrowth cells were differentiated into chondrogenic lineage with pellet culture. Chondrogenic pellets were maintained for 30 days. The quality of chondrogenic pellets was evaluated using various staining and genetic analysis of cartilage-specific markers.ResultsReprogramming was successfully done using CBMCs. CBMC-hiPSCs (n = 3) showed high pluripotency and normal karyotype. Chondrogenic pellets were generated from the outgrowth cells derived from CBMC-hiPSC EBs. The generated chondrogenic pellets showed high expression of chondrogenic genetic markers such as ACAN, COMP, COL2A1, and SOX9. The production of extracellular matrix (ECM) proteins was confirmed by safranin O, alcian blue and toluidine blue staining. Expression of collagen type II and aggrecan was detected in the accumulated ECM by immunohistological staining. Chondrogenic pellets showed low expression of fibrotic and hypertrophic cartilage marker, collagen type I and X.ConclusionsThis study reveals the potential of CBMC-hiPSCs as a promising candidate for cartilage regeneration.Electronic supplementary materialThe online version of this article (doi:10.1186/s13287-017-0477-6) contains supplementary material, which is available to authorized users.

Repair potential of nonsurgically delivered induced pluripotent stem cell-derived chondrocytes in a rat osteochondral defect model

“…We previously demonstrated the chondrogenecity of CBMC‐hiPSCs (Nam et al, ). CBMC‐hiPSCs had a comparable chondrogenic potential as MSCs.…”

“…Chondrogenesis was induced using CBMC‐hiPSCs. With this protocol, we have previously reported successful chondrogenesis using CBMC‐hiPSCs (Nam et al, ). CBMC‐hiPSC colonies had a compact morphology before use (Figure b).…”

“…It is also reported that MSCs derived from the bone marrow of elderly donors undergo inefficient chondrogenic differentiation (Baksh, Yao, & Tuan, 2007). The slow turnover rate of MSCs also limits their broad application for cartilage repair (Nam, Rim, Jung, & Ju, 2017).…”

Synthetic Poly(Vinylalcohol)‐Based Membranes for Cartilage Surgery and Repair

“…Several therapies have been or are being developed to treat this condition, including cell‐based therapies like autologous chondrocyte implantation and matrix assisted chondrocyte implantation. Stem cells have also been investigated as a cell source, including mesenchymal stem cells (MSCs) and, more recently, chondrocytes differentiated from induced pluripotent stem cells . In all cases, delivery of these cells may be by direct intra‐articular injection or seeded in a scaffold matrix which is then implanted.…”

“…Stem cells have also been investigated as a cell source, including mesenchymal stem cells (MSCs) [1] and, more recently, chondrocytes differentiated from induced pluripotent stem cells. [2] In all cases, delivery of these cells may be by direct intra-articular injection or seeded in a scaffold matrix which is then implanted. In one established procedure, the implanted cells are held in place in the defect site by a periosteal flap autograft harvested from the nearby bone.…”