Hi all,
I am working with Normal Human Epidermal Keratinocytes (NHEK) sourced from ATCC. I would like to know if any researchers here are also working with NHEK cells and can share insights, tips, or best practices.
Vishal Gaurav
Media & Supplements – Use keratinocyte growth medium (KGM) or KGM-2 with essential supplements (bovine pituitary extract, epidermal growth factor, insulin, hydrocortisone) to support proliferation. Avoid high calcium levels (>0.15 mM) unless differentiation is intended. Coating & Subculture – Preferably use collagen-coated culture surfaces to enhance adhesion. Passage cells at ~70–80% confluence using trypsin/EDTA for minimal stress. Avoid over-digestion. Seeding Density – Maintain an optimal density (~5 × 10⁴ cells/cm²) to prevent differentiation due to sparse seeding. Low Passage Number – NHEK cells have limited proliferation capacity; use early-passage cells (
NHEK cells from ATCC require optimized culture conditions to maintain viability and proper differentiation. Here are some key best practices:
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