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

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Cellular Quiescence

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1686))

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.

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Acknowledgments

H.A.C. was the Milton E. Cassel scholar of the Rita Allen Foundation. This work was funded by grants to H.A.C. from National Institute of General Medical Sciences R01 GM081686, and National Institute of General Medical Sciences R01 GM0866465. H.A.C. acknowledges a Leukemia Lymphoma Society New Idea Award, the Iris Cantor Women’s Health Center/UCLA CTSI NIH Grant UL1TR000124, Innovation Awards from the Broad Stem Cell Research Center, and a Career Development Award from the UCLA SPORE in Prostate Cancer. Research reported in this publication was supported by the National Cancer Institute of the National Institutes of Health under Award Number P50CA092131. L.D.H. acknowledges the CARE Fellows and Scholars Program. H.A.C. is a member of the Eli & Edythe Broad Center of Regenerative Medicine & Stem Cell Research, the Jonsson Comprehensive Cancer Center, the UCLA Molecular Biology Institute, and the UCLA Bioinformatics Interdepartmental Program.

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Authors and Affiliations

  1. 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

    Mithun Mitra, Linda D. Ho & Hilary A. Coller

  2. Department of Biological Chemistry, David Geffen School of Medicine, Los Angeles, CA, 90095-7329, USA

    Mithun Mitra & Hilary A. Coller

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  1. Mithun Mitra
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  2. Linda D. Ho
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  3. Hilary A. Coller
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Correspondence to Hilary A. Coller .

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  1. Texas Children’s Hospital, Baylor College of Medicine, Houston, Texas, USA

    H. Daniel Lacorazza

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Mitra, M., Ho, L.D., Coller, H.A. (2018). An In Vitro Model of Cellular Quiescence in Primary Human Dermal Fibroblasts. In: Lacorazza, H. (eds) Cellular Quiescence. Methods in Molecular Biology, vol 1686. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7371-2_2

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  • DOI: https://doi.org/10.1007/978-1-4939-7371-2_2

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