Laboratory of Viral Diseases, National Institute of Allergy and Infectious Diseases, NIH, Bethesda, Maryland 20892, USA.
J Clin Invest. 2010 Jul;120(7):2619-26. doi: 10.1172/JCI42297.
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.
原代人角质形成细胞可用于研究皮肤和黏膜上皮的许多不同疾病的发病机制。此外，它们可以在培养中形成器官型组织等效物，可作为表皮自体移植物用于伤口修复以及基因治疗的传递。然而，原代角质形成细胞在培养中具有有限的寿命，限制了它们的增殖能力和临床应用。在这里，我们报告说，用 Rho 激酶抑制剂 Y-27632 处理（源自 3 个不同解剖部位的）原代角质形成细胞可大大增加其增殖能力，并有效地实现永生化，而没有可检测到的细胞危机。更重要的是，永生化细胞表现出与原代角质形成细胞典型的特征；它们具有正常的核型和完整的 DNA 损伤反应，并且能够分化为分层上皮。这是我们所知的第一个明确的化学化合物介导有效细胞永生化的例子，这一发现可能具有广泛而深远的研究和医学应用。
