Induction and expansion of human PRRX1+ limb-bud-like mesenchymal cells from pluripotent stem cells
Induction and expansion of human PRRX1+ limb-bud-like mesenchymal cells from pluripotent stem cells
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DOI：
10.1038/s41551-021-00778-x
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发表时间：
2021-08-09
影响因子：
28.1
通讯作者：
Takarada, Takeshi
中科院分区：
文献类型：
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作者：
Yamada, Daisuke;Nakamura, Masahiro;Takarada, Takeshi
目前人类多能干细胞（hPSC）分化为软骨细胞的方案不允许扩增中间祖细胞，从而前瞻性地评估其软骨形成潜力。在这里，我们报告了一种利用 PRRX1-tdTomato 报告基因 hPSC 在确定的无异源条件下选择性诱导可扩展和个体发育定义的 PRRX1(+) 肢芽样间充质细胞的方案，并通过细胞表面标记 CD90、CD140B 和 CD82 对细胞的软骨形成潜力进行前瞻性评估。这些细胞稳定增殖并表现出进行软骨分化的潜力，形成与其 PRRX1 表达水平相称的透明软骨样组织。此外，我们还发现，源自患者的诱导性 hPSC 的肢芽样间充质细胞可用于鉴定 II 型胶原病的治疗候选者，并且我们开发了一种方法，通过将细胞铺在涂有两性离子聚合物斑点的培养皿上，产生大小均匀的透明软骨样颗粒。 PRRX1(+)肢芽样间充质细胞可以促进软骨细胞和软骨组织的大规模生产，用于药物筛选和组织工程。将人多能干细胞选择性分化为可扩展的PRRX1(+)肢芽样间充质细胞的方案可以对细胞的软骨形成潜力和透明质的形成进行前瞻性评估 软骨样颗粒。
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.A protocol for the selective differentiation of human pluripotent stem cells into expandable PRRX1(+) limb-bud-like mesenchymal cells allows for the prospective assessment of the cells' chondrogenic potential and the formation of hyaline cartilaginous-like particles.