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Split sub-confluent cultures (70-80%) 1:3 to 1:6 i.e. seeding at 0.5-1.5 x 10⁴ cells/cm² using 0.25% trypsin or trypsin/EDTA; 5% CO₂; 37°C. It is important to inactivate the trypsin with an equal volume of trypsin inhibitor. To remov
CnT-07 media This defined medium contains low calcium (0.07mM). To promote good cell attachment post trypsinisation, an 8-14 hours incubation in CnT-07 medium containing 0.2mM CaCl2 is necessary. Differentiation can be initiated by transferring a confluent monolayer from 0.07mM calcium to high calcium (1.2mM).
COCA, a murine epidermal cell line was developed by serially passaging keratinocytes from the back skin of adult C57BL/DBA mice. Culture of the COCA cells is in fully defined media without requirement for feeder layer or other coating. COCA retains its ability to differentiate and stratify in response to increased calcium concentrations thus providing an excellent experimental system for in vitro (conventional and 3D epidermal cell cultures) and in vivo (skin regeneration) skin modelling.
Mouse epidermal keratinocyes, differentiation
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