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Human keratinocyte culture. Identification and staging of epidermal cell subpopulations.

Authors
  • Staiano-Coico, L
  • Higgins, P J
  • Darzynkiewicz, Z
  • Kimmel, M
  • Gottlieb, A B
  • Pagan-Charry, I
  • Madden, M R
  • Finkelstein, J L
  • Hefton, J M
Type
Published Article
Journal
The Journal of clinical investigation
Publication Date
Feb 01, 1986
Volume
77
Issue
2
Pages
396–404
Identifiers
PMID: 2418062
Source
Medline
License
Unknown

Abstract

Stratification of human epidermal cells into multilayered sheets composed of basal and suprabasal layers (resembling the stratum germinativum and stratum spinosum of the epidermis) was studied in a dermal component-free culture system. Although no stratum corneum developed in vitro, this culture system provided a method to study early events in human keratinocyte differentiation. Multiparameter flow cytometric analysis of acridine orange-stained epidermal cells from these cultures revealed three distinct subpopulations differing in cell size, RNA content, and cell cycle kinetics. The first subpopulation was composed of small basal keratinocytes with low RNA content and a long generation time. The second subpopulation consisted of larger keratinocytes, having higher RNA content and a significantly shorter generation time. Finally, the third subpopulation contained the largest cells, which did not divide, and represent the more terminally differentiated keratinocytes. This in vitro approach provides discriminating cytochemical parameters by which the maturity of the epidermal cell sheets can be assessed prior to grafting onto human burn patients.

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