An In Vitro Model of Cellular Quiescence in Primary Human Dermal Fibroblasts

Abstract

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.

Keywords: Contact inhibition; Fibroblasts; Quiescence; Serum starvation.

Fig. 1

Methodology for generating proliferating, contact-inhibited, and serum-starved fibroblasts. A. A schematic showing the protocol for generating proliferating, 7-day contact inhibited and 7-day serum starved fibroblasts is provided. B. Images of proliferating, serum starved and contact inhibited fibroblasts are provided.