Mitra Mithun, Ho Linda D, Coller Hilary A
Department of Molecular, Cell and Developmental Biology, 5145 Terasaki Life Science Building, 610 Charles E. Young Drive E., University of California, Los Angeles, 90095-7329, USA.
Department of Biological Chemistry, David Geffen School of Medicine, Los Angeles, CA, 90095-7329, USA.
Methods Mol Biol. 2018;1686:27-47. doi: 10.1007/978-1-4939-7371-2_2.
Cellular quiescence is a reversible mode of cell cycle exit that allows cells and organisms to withstand unfavorable stress conditions. The factors that underlie the entry, exit, and maintenance of the quiescent state are crucial for understanding normal tissue development and function as well as pathological conditions such as chronic wound healing and cancer. In vitro models of quiescence have been used to understand the factors that contribute to quiescence under well-controlled experimental conditions. Here, we describe an in vitro model of quiescence that is based on neonatal human dermal fibroblasts. The fibroblasts are induced into quiescence by antiproliferative signals, contact inhibition, and serum-starvation (mitogen withdrawal). We describe the isolation of fibroblasts from skin, methods for inducing quiescence in isolated fibroblasts, and approaches to manipulate the fibroblasts in proliferating and quiescent states to determine critical regulators of quiescence.
细胞静止是一种可逆的细胞周期退出模式，它使细胞和生物体能够承受不利的应激条件。静止状态的进入、退出和维持所涉及的因素对于理解正常组织发育和功能以及诸如慢性伤口愈合和癌症等病理状况至关重要。静止的体外模型已被用于了解在严格控制的实验条件下促成静止的因素。在此，我们描述一种基于新生儿人皮肤成纤维细胞的静止体外模型。成纤维细胞通过抗增殖信号、接触抑制和血清饥饿（去除有丝分裂原）被诱导进入静止状态。我们描述了从皮肤中分离成纤维细胞的方法、在分离的成纤维细胞中诱导静止的方法，以及操纵处于增殖和静止状态的成纤维细胞以确定静止关键调节因子的方法。
