Characteristics of homogeneously small keratinocytes from newborn rat skin: possible epidermal stem cells
J. H. Pavlovitch, M. Rizk-Rabin, P. Jaffray, H. Hoehn and M. Poot Centre National de la Recherche Scientifique, Unite de Recherche Associee 583, Hopital Necker-Enfants-Malades, Paris, France. The aims of the present study were to characterize the phenotype, growth kinetics, and proliferative activat...
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Veröffentlicht in: | American Journal of Physiology: Cell Physiology 1991-12, Vol.261 (6), p.C964-C972 |
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Zusammenfassung: | J. H. Pavlovitch, M. Rizk-Rabin, P. Jaffray, H. Hoehn and M. Poot
Centre National de la Recherche Scientifique, Unite de Recherche Associee 583, Hopital Necker-Enfants-Malades, Paris, France.
The aims of the present study were to characterize the phenotype, growth
kinetics, and proliferative activation in culture of a population of poorly
differentiated homogeneously small (HS) keratinocytes. These slow-cycling
cells were separated by unit gravity sedimentation from a population of
actively proliferating basal keratinocytes in newborn rat skin. This
population (approximately 1% of the total basal keratinocytes) consisted of
extremely small cells with little cytoplasm or RNA. Their positive KL4
staining demonstrates that they were keratinocytes. HS keratinocytes did
not, however, contain epidermal calcium binding protein. Acridine orange,
bivariate Hoechst, and ethidium bromide flow cytometry of in vitro
bromodeoxyuridine-labeled cells as well as Ki67 staining showed that HS
keratinocytes were in the G0 stage of the cell cycle and did not actively
proliferate in vivo. [3H]thymidine label-retaining cells were found only in
the HS cell population, showing that HS cells may originate from a central
position in the epidermal proliferative unit. Growth of HS cells in vitro
was characterized by a delayed but progressive increase in RNA before entry
into the cell cycle. The clonogenic efficiency of HS cells in primary
culture was much less than that of larger cells. Subclones of HS cell
colonies exceeded primary colonies in their cloning efficiency and
proliferative potential, suggesting that HS cells, although normally
prevented from dividing, retain a high self-renewal capacity. They also
maintain the ability to differentiate. The results are consistent with the
concept that HS cell population may represent the epidermal-specific
progenitor cells which act as stem cells in this tissue. |
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ISSN: | 0363-6143 0002-9513 1522-1563 |
DOI: | 10.1152/ajpcell.1991.261.6.c964 |