Abstract
Studies using cultured cells allow one to dissect complex cellular mechanisms in greater detail than when studying living organisms alone. However, before cultured cells can deliver meaningful results they must accurately represent the in vivo situation. Over the last three to four decades considerable effort has been devoted to the development of culture media which improve in vitro growth and modeling accuracy. In contrast to earlier large-scale, non-specific screening of factors, in recent years the development of such media has relied increasingly on a deeper understanding of the cell’s biology and the selection of growth factors to specifically activate known biological processes. These new media now enable equal or better cell isolation and growth, using significantly simpler and less labor-intensive methodologies. Here we describe a simple method to isolate and cultivate epidermal keratinocytes from embryonic or neonatal skin on uncoated plastic using a medium specifically designed to retain epidermal keratinocyte progenitors in an undifferentiated state for improved isolation and proliferation and an alternative medium to support terminal differentiation.
The last author would like to state that she is a co-founder and the president of the Board of CELLnTEC, Advanced Cell Systems AG.
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Acknowledgments
We thank Peter Girling and Anthony Zakher for critical comments on the manuscript and their editorial contribution. This work has been supported by the Swiss National Science foundation (grants #31-59456.99 and 3100A0-107243), by the Swiss Committee for Technology and Innovation (CTI; grant 6661.3 BTS-LS), and the Martha Stiftung, Zurich, Switzerland.
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Caldelari, R., Müller, E.J. (2010). Short- and Long-Term Cultivation of Embryonic and Neonatal Murine Keratinocytes. In: Ward, A., Tosh, D. (eds) Mouse Cell Culture. Methods in Molecular Biology, vol 633. Humana Press. https://doi.org/10.1007/978-1-59745-019-5_10
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DOI: https://doi.org/10.1007/978-1-59745-019-5_10
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