title: Human induced pluripotent stem cells differentiated into chondrogenic lineage via generation of mesenchymal progenitor cells.
abstract: Human induced pluripotent stem cells (hiPSCs) exhibit pluripotency, proliferation capability, and gene expression similar to those of human embryonic stem cells (hESCs). hESCs readily form cartilaginous tissues in teratomas in vivo; despite extensive effort, however, to date no efficient method for inducing mature chondrocytes in vitro has been established. hiPSCs can also differentiate into cartilage in vivo by teratoma formation, but as with hESCs, no reliable system for in vitro chondrogenic differentiation of hiPSCs has yet been reported. Here, we examined the chondrogenic differentiation capability of hiPSCs using a multistep culture method consisting of embryoid body (EB) formation, cell outgrowth from EBs, monolayer culture of sprouted cells from EBs, and 3-dimensional pellet culture. In this culture process, the cell density of monolayer culture was critical for cell viability and subsequent differentiation capability. Monolayer-cultured cells exhibited fibroblast-like morphology and expressed markers for mesenchymal stem cells. After 2-3 weeks of pellet culture, cells in pellets exhibited a spherical morphology typical of chondrocytes and were surrounded by extracellular matrix that contained acidic proteoglycans. The expression of type II collagen and aggrecan in pellets progressively increased. Histological analysis revealed that over 70% of hiPSC-derived pellets successfully underwent chondrogenic differentiation. Using the same culture method, hESCs showed similar histological changes and gene expression, but differentiated slightly faster and more efficiently than hiPSCs. Our study demonstrates that hiPSCs can be efficiently differentiated into the chondrogenic lineage in vitro via generation of mesenchymal progenitor cells, using a simplified, multistep culture method.
authors: Department of Medicine and Clinical Science, Kyoto University Graduate School of Medicine, Kyoto, Japan.
authors: Noriaki
authors: N
authors: Koyama
authors: Masako
authors: M
authors: Miura
authors: Kazumasa
authors: K
authors: Nakao
authors: Eri
authors: E
authors: Kondo
authors: Toshihito
authors: T
authors: Fujii
authors: Daisuke
authors: D
authors: Taura
authors: Naotetsu
authors: N
authors: Kanamoto
authors: Masakatsu
authors: M
authors: Sone
authors: Akihiro
authors: A
authors: Yasoda
authors: Hiroshi
authors: H
authors: Arai
authors: Kazuhisa
authors: K
authors: Bessho
authors: Kazuwa
authors: K
authors: Nakao
mesh_terms: D000328:Adult; D000818:Animals; D000943:Antigens, Differentiation / Q000235:genetics / Q000378:metabolism; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D002478:Cells, Cultured; D019902:Chondrocytes / Q000378:metabolism*; D018920:Coculture Techniques; D058732:Embryoid Bodies / Q000502:physiology; D005260:Female; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000502:physiology*; D059630:Mesenchymal Stem Cells / Q000502:physiology*; D051379:Mice; D059467:Transcriptome
chemical_list: D000943:Antigens, Differentiation