Exploratory Study on the Stability Characteristics of Commercial HumanKeratinocytes

摘要

The purpose of this study was to determine the suitability of commerciallyavailable human keratinocytes for the initiation of vitro human epidermal organotypic models for vesicant research. Commercial normal human abdominal epithelial keratinocytes were cultured in vitro in serum free growth medium in sterile plastic culture flasks. The morphology, ultrastructure, growth and viability of the developing serially cultured keratinocyte cell lines were observed intermittently, and documented by phase contrast and transmission electron microscopy. First passage keratinocytes appeared normal. However, many contained paranuclear vacuole and cytoplasmic inclusions. Viability, as determined by the dye exclusion method, dropped from 94% in the second passage to 62% in the fifth passage. A total of 17.73 population doublings were obtained between the first and fifth passage (exclusive of the primary passage). Data obtained showed that senescence and differentiation occurred in the third passage. No evidence was observed for the presence of stem cells in the cell lines. This study established that commercial abdominal keratinocyte cultures were useful through the second passage for short-term experimental assays, but not for long-term epidermal organotypic modeling. Keywords: RA V; Human abdominal keratinocytes; Tissue culture; Differentiation; Growth; Morphology; Senescence; Microscopy; In vitro models; Epidermis. (kt)