TY - CHAP
T1 - A Method for In Vitro Fabrication of Hybrid Bone/Cartilage Tissue Using Mouse Induced Pluripotent Stem Cells
AU - Limraksasin, Phoonsuk
AU - Egusa, Hiroshi
N1 - Funding Information:
This work was funded by the Japan Society for the Promotion of Science (Grants-in Aid for Scientific Research B: 19H03840 and Challenging Exploratory Research: 16K15797).
Publisher Copyright:
© 2021, Springer Science+Business Media, LLC.
PY - 2022
Y1 - 2022
N2 - In the developing embryo, bone and cartilage share the same progenitors. However, osteo-chondrogenic induction of mouse induced pluripotent stem cells (iPSCs) remains difficult. Here we describe a protocol to guide iPSCs to differentiate into osteochondral cells that form hybrid bone/cartilage constructs in vitro. Single mouse iPSCs are first reaggregated in ultra-low-attachment micro-space culture plates. At day 12, iPSC spheres are subjected to shaking culture and maintained in an osteogenic induction medium for 31 days (Os induction). In another condition, the osteogenic induction medium is replaced by chondrogenic induction medium at day 22 and maintained for a further 21 days (Os-Chon induction). Os induction produced robust mineralization and some cartilage-like tissue, whereas Os-Chon induction resulted in partial mineralization and a large area of cartilage tissue.
AB - In the developing embryo, bone and cartilage share the same progenitors. However, osteo-chondrogenic induction of mouse induced pluripotent stem cells (iPSCs) remains difficult. Here we describe a protocol to guide iPSCs to differentiate into osteochondral cells that form hybrid bone/cartilage constructs in vitro. Single mouse iPSCs are first reaggregated in ultra-low-attachment micro-space culture plates. At day 12, iPSC spheres are subjected to shaking culture and maintained in an osteogenic induction medium for 31 days (Os induction). In another condition, the osteogenic induction medium is replaced by chondrogenic induction medium at day 22 and maintained for a further 21 days (Os-Chon induction). Os induction produced robust mineralization and some cartilage-like tissue, whereas Os-Chon induction resulted in partial mineralization and a large area of cartilage tissue.
KW - Hybrid bone/cartilage tissue
KW - Induced pluripotent stem cells
KW - Osteo-chondrogenic differentiation
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U2 - 10.1007/7651_2021_361
DO - 10.1007/7651_2021_361
M3 - Chapter
C2 - 33755902
AN - SCOPUS:85132454074
T3 - Methods in Molecular Biology
SP - 509
EP - 520
BT - Methods in Molecular Biology
PB - Humana Press Inc.
ER -