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Flow Cytometric Sorting and Keratin Analysis of Human Epidermal Basal Cells

1987; Wiley; Volume: 14; Issue: 1 Linguagem: Inglês

10.1111/j.1346-8138.1987.tb02986.x

1346-8138

Marie‐Jeanne Staquet, Jean‐Jacques Lawrence, Jacques Albert, J Thivolet,

melanin and skin pigmentation

Abstract A prerequisite condition for the elucidation of the expression of keratin subunits in human basal cells is obtaining highly purified basal cell fractions. A method has therefore been developed for the isolation of the basal cell sub‐population from normal human epidermis at very high purity. Epidermal cell suspensions obtained by trypsinization at 4°C were first enriched in basal cells by selective attachment to collagen. The cells were then stained in suspension with KLI monoclonal antikeratin antibody which had been shown to be reactive with suprabasal cells. The suprabasal cells contaminating the suspensions were then eliminated by selective flow cytometry sorting and basal keratinocyte populations of 99.5% purity were obtained. Analysis of the keratin composition of these cells showed that the large majority of basal cells contained two keratins, one from each keratin subfamily: type I acidic, 50 Kd [keratin no 14 as defined by Moll et al. (1)] and type II basic, 58 Kd [no 5]. Nevertheless, a small subpopulation of basal cells (2%) was shown to express, in addition to the 50/58 Kd subunits, the 56.5/65–67 Kd keratins (nos 10, 1–2) which are regarded as markers for cells committed to terminal differentiation.