Y-27632 simplifies the isolation procedure of human primary epidermal cells by selectively blocking focal adhesion of dermal cells.
Y-27632 simplifies the isolation procedure of human primary epidermal cells by selectively blocking focal adhesion of dermal cells.
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Y-27632 通过选择性阻断真皮细胞的粘斑，简化了人原代表皮细胞的分离程序。
DOI：
10.1002/term.2526
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发表时间：
2018
期刊：
影响因子：
--
通讯作者：
Wu Xunwei
中科院分区：
文献类型：
--
作者：
Wen Jie;Zu Tingjian;Zhou Qian;Leng Xue;Wu Xunwei
原代皮肤表皮细胞的分离和体外扩增培养已广泛应用于实验室研究和临床应用。传统的方法涉及顺序酶消化成人组织已经给出了低的细胞回收率和降低细胞活力。我们在此报告了一种从皮肤组织中分离人原发性表皮祖细胞的先进方法，其中包括Rho激酶抑制剂Y‐27632。与传统协议相比，当前协议具有简单、方便、快速等特点；此外，它还能提高表达整合素的上皮干细胞的产量。此外，我们的新方法不需要分离表皮和真皮，因为培养基选择性地阻断了真皮细胞的黏附和生长。重要的是，从这种方法分离的细胞可以保持其再生潜力，并在移植到裸鼠身上后快速在体内重建成熟的人类皮肤。简而言之，我们在这里描述了一种简单（一步）和无血清的方法，用于从成人组织中分离原代表皮干细胞。分离的细胞可广泛用于实验室研究和临床应用，特别是在组织工程和再生领域。
Primary skin epidermal cells isolation and in vitro expansion culture have been widely used for laboratory research and clinical applications. The conventional methods involving sequential enzymatic digestion of adult tissues have given low cell recovery rate and reduced cell viability. We report here an advanced method for human primary epidermal progenitor cells isolation from skin tissues including the Rho kinase inhibitor Y‐27632. Compared with traditional protocols, the current protocol is simple, easy, and faster; moreover, it gives a greater yield of integrin‐expressing epithelial stem cells. In addition, our new methodology does not require a separation of epidermis from dermis because the medium selectively blocks focal adhesion and growth of dermal cells. Importantly, the cells isolated from this method can maintain their regeneration potential and quickly reconstitute a mature human skin in vivo after grafting onto nude mice. In brief, we describe here a simple (one step) and serum‐free method for isolating primary epidermal stem cells from adult tissues. The isolated cells may be widely used for both laboratory studies and clinical application, especially in the field of tissue engineering and regeneration.