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.
Yorkshire Regional Tissue Bank, Pinderfields General Hospital, Wakefield, UK.
J T
JT
Daniels
I R
IR
Harris
J N
JN
Kearney
E
E
Ingham
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