ArticleMethods in molecular biology (Clifton, N.J.)2025
Expansion of Human Epidermal Keratinocytes in Co-culture with Human Feeder Cells.
Article in Methods in molecular biology (Clifton, N.J.), 2025. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.
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Abstract
The co-culture of human epidermal keratinocytes with mouse 3T3-J2 feeder cells, a method developed by Green and colleagues, has been widely used since the early 1980s to generate skin autografts. Additionally, co-culture with 3T3-J2 cells has been a crucial tool in skin stem cell biology, as it facilitates the evaluation of self-renewal capacity and differentiation of epidermal stem cells. This chapter presents a recent enhancement of the Green method, aimed at further promoting the expansion of human epidermal keratinocytes by utilizing a small molecule inhibitor of TGF-β signaling. This new protocol also enables the rapid expansion of human epidermal keratinocytes in co-culture with human feeder cells, including human dermal fibroblasts and human preadipocytes-two key alternatives to 3T3-J2 cells-with the long-term goal of developing customized Xeno-free skin autografts.
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