Mouse models have been used to study the physiology and pathogenesis of the skin. However, propagation of mouse primary epidermal keratinocytes remains challenging. In this chapter, we introduce a newly developed protocol that enables long-term expansion of p63+ mouse epidermal keratinocytes in low-Ca2+ media without the need of progenitor cell purification steps or support by a feeder cell layer. Pharmacological inhibition of TGF-β signaling in crude preparations of mouse epidermis robustly increases proliferative capacity of p63+ epidermal progenitor cells while preserving their ability to differentiate. Suppression of TGF-β signaling also permits p63+ epidermal keratinocytes to form macroscopically large clones in 3T3-J2 feeder cell co-culture. This simple and efficient approach will facilitate the use of mouse models by providing p63+ primary epidermal keratinocytes in quantity.
CITATION STYLE
Pinto, F., Suzuki, D., & Senoo, M. (2019). Long-term expansion of mouse primary epidermal keratinocytes using a TGF-β signaling inhibitor. In Methods in Molecular Biology (Vol. 1993, pp. 47–59). Humana Press Inc. https://doi.org/10.1007/978-1-4939-9473-1_4
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