Calcium: a crucial consideration in serum-free keratinocyte culture.
This investigation was conducted when previously repeatable experimental data became impossible to reproduce when using keratinocytes cultured in serum-free medium. Differences in calcium molarity between batches of medium were identified as a source of variation in cultured keratinocyte populations. The susceptibility of cultured keratinocytes to even small alterations in calcium molarity has been demonstrated. 2 regular medium batches were compared with a special preparation of medium, devoid of calcium chloride then supplemented with a known concentration of calcium ions. Culture progress in each medium was assessed by: morphological observation, % cells expressing involucrin and proliferating cell nuclear antigen, cell attachment, growth rate and colony forming efficiency. In order to control the phenotype of cultured keratinocytes, in a reproducible system, it is recommended that serum-free keratinocytes medium is purchased with the omission of calcium chloride. Supplementation of this medium may then be made by the investigator to suit individual culture requirements.
Daniels
J T
JT
Yorkshire Regional Tissue Bank, Pinderfields General Hospital, Wakefield, UK.
Harris
I R
IR
Kearney
J N
JN
Ingham
E
E
D002118:Calcium; D002455:Cell Division; D002478:Cells, Cultured; D016895:Culture Media, Serum-Free; D006801:Humans; D007150:Immunohistochemistry; D015603:Keratinocytes / Q000166:cytology*; D018809:Proliferating Cell Nuclear Antigen; D011498:Protein Precursors
D016895:Culture Media, Serum-Free; D018809:Proliferating Cell Nuclear Antigen; D011498:Protein Precursors; C032253:involucrin; D002118:Calcium