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Yes primary mouse keratinocytes grow well in this media when you reduce the CaCl2 concentration to 0.025 mM - 0.05 mM CaCl2 (optimal Ca concentration for primary human keratinocytes is 0.06 mM).
If you are growing our NHEK in PromoCell Keratinocyte Growth Medium 2 (C-20011) or Growth Medium 3 (C-20021) you don't need any feeder cells. The cells will grow as a monolayer in conventional tissue culture flasks.
The optimal calcium concentration for both proliferation and differentiation of keratinocytes depends on the species and also on the media formulation. To keep primary human keratinocytes in the proliferative status concentrations between 0.03 and 0.15 mM (PromoCell Keratinocyte Growth Medium 2: 0.06 mM) are generally used. Increasing the calcium above 1 mM will induce terminal differentiation and lead to the loss of proliferative activity.
Yes; the NHEK-GM2 (C-12001; C-12003; C-12005; C-12006) also grow in PromoCell Keratinocyte Growth Medium 3 (C-20021). Using the protocol with the fixed intervals; they grow slightly faster than in Keratinocyte GM2 and proliferate for > 15 PDs.
Conversely; NHEK-GM3 (C-12011; C-12013; C-12015; C-12016) also grow in the existing Keratinocyte Growth Medium 2 (C-20011). When using the classical subcultivation protocol (density > 70% - 90%); they grow slightly slower compared to Keratinocyte GM3; but also reach > 15 PDs.
PromoCell's Normal Human Epidermal Keratinocytes (NHEK) from juvenile donors are isolated from both the epidermis of the outer and mucosal (i.e. inner) layers of the foreskin. Thus; each vial contains a mixture of mucosal and cutaneous keratinocytes.
PromoCell is using a pan-cytokeratin antibody rather than a specific type of cytokeratin for the quality control of our keratinocytes.