Growth and differentiation of mouse epidermal cells in culture: effects of extracellular calcium.
The pattern of proliferation and differentiation in cultured mouse epidermal cells in markedly altered by modifying the ionic calcium concentration in the culture medium. When medium calcium is lowered from 1.44 mM to 0.05-0.1 mM, keratinocytes proliferate rapidly with a high growth fraction, do not stratify, but continue to synthesize keratin. The cells grow as a monolayer for several months and can be subcultured in low Ca++ medium. Ultrastructural examination of cells cultured under low Ca++ conditions reveals widened intercellular spaces with an absence of desmosomes. Microvilli are numerous, and tonofilaments and cellular organelles are organized perinuclearly. Epidermal cells growing as a monolayer in low Ca++ can be identified to terminally differentiate by adding calcium to the level normally found in the culture medium. Contact between cells occurs rapidly and desmosomes form within 2 hours. The cells stratify in 1-2 days and terminally differentiate in 3-4 days. After Ca++ addition, DNA synthesis decreases after a lag of 5-10 hours and is totally inhibited within 36 hours. In contrast, RNA and protein synthesis continue at 40-50% of the control level at Day 3, a time when many cells are detaching from the culture dish. Keratin synthesis is unaffected by the Ca++ switch. Manipulation of epidermal proliferation and differentiation by altering extracellular calcium levels should enhance the usefulness of epidermal cell cultures in the study of differentiation and carcinogenesis.
D000818:Animals; D002118:Calcium / Q000494:pharmacology*; D002454:Cell Differentiation / Q000187:drug effects; D002455:Cell Division / Q000187:drug effects; D002478:Cells, Cultured; D004247:DNA / Q000096:biosynthesis; D004817:Epidermis / Q000502:physiology*; D051379:Mice; D014176:Protein Biosynthesis; D013997:Time Factors
D004247:DNA; D002118:Calcium