Human keratinocytes are efficiently immortalized by a Rho kinase inhibitor
Human keratinocytes are efficiently immortalized by a Rho kinase inhibitor
复制标题
DOI：
10.1172/jci42297
复制
发表时间：
2010-07-01
影响因子：
15.9
通讯作者：
McBride, Alison A.
中科院分区：
文献类型：
--
作者：
Chapman, Sandra;Liu, Xuefeng;McBride, Alison A.
原代人角质形成细胞可用于研究皮肤和粘膜上皮的许多不同疾病的发病机制。此外，它们可以在培养物中形成器官型组织等同物，可用作表皮自体移植物，用于伤口修复以及基因治疗的实施。然而，原代角质形成细胞在培养物中的寿命有限，限制了其增殖能力和临床应用。在这里，我们报告用 Rho 激酶抑制剂 Y-27632 处理原代角质形成细胞（源自 3 个不同的解剖部位），大大提高了其增殖能力，并导致有效的永生化，而没有检测到细胞危机。更重要的是，永生化细胞表现出原代角质形成细胞的典型特征。它们具有正常的核型和完整的DNA损伤反应，并且能够分化成复层上皮。这是我们对介导有效细胞永生化的特定化合物的了解的第一个例子，这一发现可能具有广泛而深刻的研究和医学应用。
Primary human keratinocytes are useful for studying the pathogenesis of many different diseases of the cutaneous and mucosal epithelia. In addition, they can form organotypic tissue equivalents in culture that can be used as epidermal autografts for wound repair as well as for the delivery of gene therapy. However, primary keratinocytes have a finite lifespan in culture that limits their proliferative capacity and clinical use. Here, we report that treatment of primary keratinocytes (originating from 3 different anatomical sites) with Y-27632, a Rho kinase inhibitor, greatly increased their proliferative capacity and resulted in efficient immortalization without detectable cell crisis. More importantly, the immortalized cells displayed characteristics typical of primary keratinocytes; they had a normal karyotype and an intact DNA damage response and were able to differentiate into a stratified epithelium. This is the first example to our knowledge of a defined chemical compound mediating efficient cell immortalization, and this finding could have wide-ranging and profound investigational and medical applications.