TY - JOUR
T1 - Induction and expansion of human PRRX1+ limb-bud-like mesenchymal cells from pluripotent stem cells
AU - Yamada, Daisuke
AU - Nakamura, Masahiro
AU - Takao, Tomoka
AU - Takihira, Shota
AU - Yoshida, Aki
AU - Kawai, Shunsuke
AU - Miura, Akihiro
AU - Ming, Lu
AU - Yoshitomi, Hiroyuki
AU - Gozu, Mai
AU - Okamoto, Kumi
AU - Hojo, Hironori
AU - Kusaka, Naoyuki
AU - Iwai, Ryosuke
AU - Nakata, Eiji
AU - Ozaki, Toshifumi
AU - Toguchida, Junya
AU - Takarada, Takeshi
N1 - Funding Information:
We thank N. Tsumaki for providing iPS cell lines derived from patients with COL2pathy (ACGII-1 and HCG-1); M. Hada and M. Nishio for technical assistance with feeder-free cultures; A. Hirao for providing chemical libraries; and K. Sekiguchi, S. Nagata, S. Tamaki, Y. Jin and C. Alev for their invaluable comments and advice. Preparation of slides and staining was supported by the Central Research Laboratory, Okayama University Medical School. We thank the members of the Department of Animal Resources, Advanced Science Research Center and Okayama University for maintaining the mice. This research was supported by Grants-in-aid for Scientific Research from the Japan Society for the Promotion of Science (grant no. 17H04399 to T. Takarada), AMED (grant nos 18bm0704024h0001 and 20bm0404064h0001 to T. Takarada), the Acceleration Program for Intractable Disease Research Utilizing Disease Specific iPS Cells (AMED) to J.T. and the Cooperative Research Program (Joint Usage/Research Center program) of the Institute for Frontier Life and Medical Sciences, Kyoto University to T. Takarada and J.T. These funders had no role in the study design, data collection and analysis, decision to publish or preparation of the manuscript.
Publisher Copyright:
© 2021, The Author(s), under exclusive licence to Springer Nature Limited.
PY - 2021/8
Y1 - 2021/8
N2 - Current protocols for the differentiation of human pluripotent stem cells (hPSCs) into chondrocytes do not allow for the expansion of intermediate progenitors so as to prospectively assess their chondrogenic potential. Here we report a protocol that leverages PRRX1–tdTomato reporter hPSCs for the selective induction of expandable and ontogenetically defined PRRX1+ limb-bud-like mesenchymal cells under defined xeno-free conditions, and the prospective assessment of the cells’ chondrogenic potential via the cell-surface markers CD90, CD140B and CD82. The cells, which proliferated stably and exhibited the potential to undergo chondrogenic differentiation, formed hyaline cartilaginous-like tissue commensurate to their PRRX1-expression levels. Moreover, we show that limb-bud-like mesenchymal cells derived from patient-derived induced hPSCs can be used to identify therapeutic candidates for type II collagenopathy and we developed a method to generate uniformly sized hyaline cartilaginous-like particles by plating the cells on culture dishes coated with spots of a zwitterionic polymer. PRRX1+ limb-bud-like mesenchymal cells could facilitate the mass production of chondrocytes and cartilaginous tissues for applications in drug screening and tissue engineering.
AB - Current protocols for the differentiation of human pluripotent stem cells (hPSCs) into chondrocytes do not allow for the expansion of intermediate progenitors so as to prospectively assess their chondrogenic potential. Here we report a protocol that leverages PRRX1–tdTomato reporter hPSCs for the selective induction of expandable and ontogenetically defined PRRX1+ limb-bud-like mesenchymal cells under defined xeno-free conditions, and the prospective assessment of the cells’ chondrogenic potential via the cell-surface markers CD90, CD140B and CD82. The cells, which proliferated stably and exhibited the potential to undergo chondrogenic differentiation, formed hyaline cartilaginous-like tissue commensurate to their PRRX1-expression levels. Moreover, we show that limb-bud-like mesenchymal cells derived from patient-derived induced hPSCs can be used to identify therapeutic candidates for type II collagenopathy and we developed a method to generate uniformly sized hyaline cartilaginous-like particles by plating the cells on culture dishes coated with spots of a zwitterionic polymer. PRRX1+ limb-bud-like mesenchymal cells could facilitate the mass production of chondrocytes and cartilaginous tissues for applications in drug screening and tissue engineering.
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U2 - 10.1038/s41551-021-00778-x
DO - 10.1038/s41551-021-00778-x
M3 - Article
C2 - 34373601
AN - SCOPUS:85112652649
SN - 2157-846X
VL - 5
SP - 926
EP - 940
JO - Nature Biomedical Engineering
JF - Nature Biomedical Engineering
IS - 8
ER -