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Promotion of Human Epidermal Keratinocyte Expansion in Feeder Cell Co-culture

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Part of the Methods in Molecular Biology book series (MIMB, volume 1993)

Abstract

Co-culture of human epidermal keratinocytes with mouse 3T3-J2 feeder cells, developed by Green and colleagues, has been used worldwide to generate skin autografts since the early 1980s. In addition, co-culture with 3T3-J2 cells has served as a fundamental tool in skin stem cell biology as it allows the evaluation of self-renewal capacity of epidermal stem cells. This chapter describes a recent improvement in the Green method to promote further the expansion of human epidermal keratinocytes utilizing a small-molecule inhibitor of TGF-β signaling. This new protocol enables more rapid expansion of human epidermal keratinocytes in co-culture with not only 3T3-J2 cells but also other feeder cells including human dermal fibroblasts and human preadipocytes, two major alternatives to 3T3-J2 cells, with a long-term goal of developing customized skin autografts.

Key words

Human keratinocytes Skin transplantation Feeder cell co-culture TGF-β signaling Small-molecule inhibitor 
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Notes

Acknowledgments

The authors thank Jeff Holcombe for proofreading of the manuscript. This study was supported by an R01AR066755 grant from the National Institute of Arthritis and Musculoskeletal and Skin Diseases of the National Institute of Health to M.S.

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© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Department of Molecular and Cell BiologyBoston University Henry M. Goldman School of Dental MedicineBostonUSA

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