The effect of vitamin A acid on the culture of guinea pig ear keratinocytes was investigated. Use of this agent and treatment of the plastic dishes with collagen solution facilitated attachment and growth. Vitamin A acid elicited a wave of DNA synthesis which was studied by incorporation of tritiate
Effect of vitamin A acid on cyclic nucleotides of cultured keratinocytes
β Scribed by David I. Wilkinson; Elaine K. Orenberg
- Publisher
- Springer-Verlag
- Year
- 1980
- Tongue
- English
- Weight
- 397 KB
- Volume
- 267
- Category
- Article
- ISSN
- 0340-3696
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β¦ Synopsis
Primary cultures of guinea pig ear keratinocytes were treated with vitamin A acid at plating, or at 14 h or 14 days after plating. The intracellular content of the cyclic nucleotides cAMP and cGMP was determined by radioimmunoassay at intervals during a period of 50 h after treatment. When added at plating, vitamin A acid produced a wave of DNA synthesis and increase in DNA which was at maximum between 30 and 40 h after plating, and coincided with decreased cAMP levels. This may represent a subpopulation of keratinocytes in S phase. Treatment with vitamin A acid at 14 h or 14 days after plating resulted in an immediate but temporary fall in cAMP and cGMP, and a wave of thymidine uptake but no increase in DNA per dish. Thus, a single treatment with vitamin A acid is mitogenic only when applied at plating. At other times, it can cause changes in cyclic nucleotide content without any observable cell proliferation.
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Cultured human keratinocytes were incubated in the presence of glycolipids (GM1, asialoGM1, glucosylceramide and galactosyiceramide) added to the medium at a concentration of 10-SM. All of them became associated with the cells, but in different amounts depending on the type of glycolipid and on the