Isolation and short-term culture of primary keratinocytes, hair follicle populations and dermal cells from newborn mice and keratinocytes from adult mice for in vitro analysis and for grafting to immunodeficient mice
Top Cited Papers
- 10 April 2008
- journal article
- research article
- Published by Springer Science and Business Media LLC in Nature Protocols
- Vol. 3 (5), 799-810
- https://doi.org/10.1038/nprot.2008.50
Abstract
Protocols for preparing and culturing primary keratinocytes from newborn and adult mouse epidermis have evolved over the past 35 years. This protocol is now routinely applied to mice of various genetic backgrounds for in vitro studies of signaling pathways in differentiation and cell transformation, and for assessing the in vivo phenotype of altered keratinocytes in grafts of cells on immunodeficient mice. Crucial in the development and application of the procedure was the observation that keratinocytes proliferate in media of low calcium concentration, but rapidly commit to differentiation at calcium concentrations >0.07 mM after the initial attachment period. Preparing primary keratinocytes from ten newborn mice requires 2–3 h of hands-on time. Related procedures are also provided: preparing immature hair follicle buds, developing dermal hair follicles and fibroblasts from newborn mice, preparing primary keratinocytes from adult mice and grafting cell mixtures on athymic nude mice.Keywords
This publication has 19 references indexed in Scilit:
- Repair characteristics and differentiation propensity of long-term cultures of epidermal keratinocytes derived from normal and NER-deficient miceDNA Repair, 2005
- Long-Term Culture of Murine Epidermal KeratinocytesJournal of Investigative Dermatology, 1999
- Formation of the Epidermal Calcium Gradient Coincides with Key Milestones of Barrier Ontogenesis in the RodentJournal of Investigative Dermatology, 1998
- [1] Isolation and utilization of epidermal keratinocytes for oncogene researchMethods in enzymology, 1995
- Expression of murine epidermal differentiation markers is tightly regulated by restricted extracellular calcium concentrations in vitro.The Journal of cell biology, 1989
- Spontaneous establishment and characterization of mouse keratinocyte cell lines in serum-free mediumIn Vitro Cellular & Developmental Biology, 1988
- BALB and kirsten murine sarcoma viruses alter growth and differentiation of EGF-dependent BALB/c mouse epidermal keratinocyte linesCell, 1983
- Clonal Growth of Mouse Epidermal Cells in Medium with Reduced Calcium ConcentrationJournal of Investigative Dermatology, 1981
- Calcium regulation of growth and differentiation of mouse epidermal cells in cultureCell, 1980
- Mouse epidermal cell cultures: II. Isolation, characterization and cultivation of epidermal cells from perinatal mouse skinExperimental Cell Research, 1975