Impacts of manipulating cell sorting on in vitro hair follicle regeneration
Impacts of manipulating cell sorting on in vitro hair follicle regeneration
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DOI：
10.1016/j.jbiosc.2022.09.004
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发表时间：
2022-12-03
影响因子：
2.8
通讯作者：
Fukuda, Junji
中科院分区：
文献类型：
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作者：
Kageyama, Tatsuto;Anakama, Riki;Fukuda, Junji
毛囊形态发生是由上皮-间充质相互作用触发的。已经开发了几种方法用于使用上皮细胞和间充质细胞制备毛囊类器官;然而，目前对体外自发组织过程与毛发再生的相关性的理解是有限的。在本研究中，我们使用Y27632，一种rho相关激酶抑制剂，研究细胞分选操作对体外毛发再生的影响。将分离的毛囊诱导型上皮细胞和间充质细胞在96孔板或聚二甲基硅氧烷微阵列板中的含Y27632的培养基中培养。我们发现，Y27632补充调制的空间分布的上皮细胞和间充质细胞从哑铃形核壳配置通过自发组织过程。在Y27632处理的核壳状聚集体中出现具有典型形态特征的新毛囊，并且毛干在体外以约100%的效率发芽。基因芯片分析和通路抑制实验表明，磷脂酰肌醇-3-激酶/蛋白激酶B-和Ras-信号通路参与了Y27632处理的毛囊类器官中的毛发样发芽。我们的发现增强了对毛囊器官发生和毛囊类器官发育的理解。(c)2022年，生物技术学会，日本。All rights reserved.
Hair follicle morphogenesis is triggered by epithelialemesenchymal interactions. Several approaches have been developed for preparing hair follicle organoids using epithelial and mesenchymal cells; however, the current understanding of the relevance of in vitro spontaneous organization processes to hair regeneration is limited. In the present study, we used Y27632, a rho-associated kinase inhibitor, to investigate the effects of manipulation of cell sorting on hair regeneration in vitro. Dissociated hair follicle-inducible epithelial and mesenchymal cells were cultured in Y27632containing media in 96-well plates or polydimethylsiloxane microarray plates. We found that Y27632 supplementation modulated the spatial distribution of epithelial and mesenchymal cells from a dumbbell shape to a coreeshell configuration via a spontaneous organization process. New hair follicles with typical morphological features emerged in the Y27632-treated core-shell-shaped aggregates, and hair shafts sprouted with approximately 100% efficiency in vitro. Gene chip analysis and pathway-inhibition experiments revealed that the phosphatidylinositol-3-kinase/protein kinase B- and Ras-signaling pathways were involved in hair-like sprouting in the Y27632-treated hair follicle organoids. Our findings enhance the understanding of hair follicle organogenesis and the development of hair follicle organoids. (c) 2022, The Society for Biotechnology, Japan. All rights reserved.