Generation of scaffoldless hyaline cartilaginous tissue from human iPSCs.
Defects in articular cartilage ultimately result in loss of joint function. Repairing cartilage defects requires cell sources. We developed an approach to generate scaffoldless hyaline cartilage from human induced pluripotent stem cells (hiPSCs). We initially generated an hiPSC line that specifically expressed GFP in cartilage when teratoma was formed. We optimized the culture conditions and found BMP2, transforming growth factor β1 (TGF-β1), and GDF5 critical for GFP expression and thus chondrogenic differentiation of the hiPSCs. The subsequent use of scaffoldless suspension culture contributed to purification, producing homogenous cartilaginous particles. Subcutaneous transplantation of the hiPSC-derived particles generated hyaline cartilage that expressed type II collagen, but not type I collagen, in immunodeficiency mice. Transplantation of the particles into joint surface defects in immunodeficiency rats and immunosuppressed mini-pigs indicated that neocartilage survived and had potential for integration into native cartilage. The immunodeficiency mice and rats suffered from neither tumors nor ectopic tissue formation. The hiPSC-derived cartilaginous particles constitute a viable cell source for regenerating cartilage defects.
Cell Induction and Regulation Field, Department of Cell Growth and Differentiation, Center for iPS Cell Research and Application, Kyoto University, Kyoto 606-8507, Japan.
Akihiro
A
Yamashita
Cell Induction and Regulation Field, Department of Cell Growth and Differentiation, Center for iPS Cell Research and Application, Kyoto University, Kyoto 606-8507, Japan.
Miho
M
Morioka
Cell Induction and Regulation Field, Department of Cell Growth and Differentiation, Center for iPS Cell Research and Application, Kyoto University, Kyoto 606-8507, Japan.
Yasuhito
Y
Yahara
Cell Induction and Regulation Field, Department of Cell Growth and Differentiation, Center for iPS Cell Research and Application, Kyoto University, Kyoto 606-8507, Japan.
Minoru
M
Okada
Cell Induction and Regulation Field, Department of Cell Growth and Differentiation, Center for iPS Cell Research and Application, Kyoto University, Kyoto 606-8507, Japan; Department of Orthopaedic Surgery, Graduate School of Medicine, Kyoto University, Kyoto 606-8507, Japan.
Tomohito
T
Kobayashi
Department of Orthopaedic Surgery, Graduate School of Medicine, Kyoto University, Kyoto 606-8507, Japan.
Shinichi
S
Kuriyama
Department of Orthopaedic Surgery, Graduate School of Medicine, Kyoto University, Kyoto 606-8507, Japan.
Shuichi
S
Matsuda
Cell Induction and Regulation Field, Department of Cell Growth and Differentiation, Center for iPS Cell Research and Application, Kyoto University, Kyoto 606-8507, Japan; Japan Science and Technology Agency, CREST, Tokyo 102-0075, Japan. Electronic address: ntsumaki@cira.kyoto-u.ac.jp.
Noriyuki
N
Tsumaki
D000818:Animals; D015415:Biomarkers; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D002460:Cell Line; D002471:Cell Transformation, Neoplastic; D019902:Chondrocytes / Q000166:cytology; D020219:Chondrogenesis; D015870:Gene Expression; D055879:Gene Knock-In Techniques; D017930:Genes, Reporter; D006801:Humans; D051457:Hyaline Cartilage / Q000166:cytology* / Q000378:metabolism*; D057026:Induced Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism*; D051379:Mice; D016513:Mice, SCID; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D033581:Stem Cell Transplantation; D013552:Swine; D013556:Swine, Miniature; D019076:Transgenes
D015415:Biomarkers; D011993:Recombinant Fusion Proteins