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Methods Mol Biol. 2019;1993:15-31. doi: 10.1007/978-1-4939-9473-1_2.

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

Author information

1
Department of Molecular and Cell Biology, Boston University Henry M. Goldman School of Dental Medicine, Boston, MA, USA.
2
Department of Molecular and Cell Biology, Boston University Henry M. Goldman School of Dental Medicine, Boston, MA, USA. msenoo@bu.edu.

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.

KEYWORDS:

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

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