Cell subpopulations within proliferative and differentiating compartments of epidermis

Pavlovitch, J. H.; Rizk-Rabin, Marthe; M, Gervaise; Metezeau, Philippe; Grünwald, Didier

doi:10.1152/ajpcell.1989.256.5.c977

Cited by 13 publications

(14 citation statements)

References 20 publications

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“…It has been shown that a feature of stem cell populations, given their primitive state, is that they are homogeneously small, blast-like cells with a high N͞C ratio (2,(25)(26)(27). Initial flowcytometric analysis revealed that epidermal SPs exhibited homogeneously low intracellular complexity (i.e., low SSC) and formed a cluster in the low forward-scatter region (Fig.…”

Section: Resultsmentioning

confidence: 99%

Side population in adult murine epidermis exhibits phenotypic and functional characteristics of keratinocyte stem cells

Redvers

Kaur

2006

Proc. Natl. Acad. Sci. U.S.A.

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Based on functional studies in the bone marrow, it has been suggested that the ability to efflux Hoechst 33342 may represent a universal stem cell trait. In this phenotypic and functional characterization of the Hoechst side population (SP) in adult murine epidermis, we demonstrate that these cells are a rare subset of the keratinocyte stem cell-enriched ␣6 bri CD71 dim fraction comprising SSC low ͞K14 ؉ ͞CD34 ؊ ͞Oil red O ؊ ͞c-kit ؊ ͞CD45 ؊ keratinocytes. Epidermal SPs have the smallest cell and nuclear size but exhibit the highest nuclear-to-cytoplasmic ratio of any fraction examined, consistent with a primitive cell type. Although SPs demonstrated poor cumulative in vitro proliferative output, they exhibited sustained epidermal tissue-regenerative activity in vivo compared with unfractionated and non-SP cells. Collectively, these results indicate that the epidermal SP contains the most potent keratinocyte stem cell population in skin epithelium.skin ͉ self-renewal ͉ tissue regeneration ͉ cell cycle ͉ proliferation

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Section: Resultsmentioning

confidence: 99%

Side population in adult murine epidermis exhibits phenotypic and functional characteristics of keratinocyte stem cells

Redvers

Kaur

2006

Proc. Natl. Acad. Sci. U.S.A.

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“…Keratinocytes were separated in a specially designed sedimentation chamber (Tulp et al, 1980) according to a procedure described previously (Pavlovitch et al, 1989). Cells were either cultured (see below) or stored at -20°C in MEM supplemented with 10% FBS and 10% dimethylsulfoxide.…”

Section: Unit-gravity Sedimentationmentioning

confidence: 99%

“…To elucidate the complex relationships between cell size, cell differentiation, and proliferative capacity, we separated keratinocytes from the basal layer of the rat epidermis in a unit-gravity separation chamber (Pavlovitch et al, 1989). The populations of different cell sizes thus obtained were either directly analysed as to RNA and DNA content, which reveals the extent of cell differentiation, or brought into culture to assess their proliferative capacity.…”

mentioning

confidence: 99%

Cell size and RNA content correlate with cell differentiation and proliferative capacity of rat keratinocytes

Poot

Rizk-Rabin

Hoehn

et al. 1990

Journal Cellular Physiology

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Keratinocytes from rat skin were separated according to their size in a specially designed unit-gravity sedimentation chamber. The fractions obtained with this technique showed clear morphological differences, and analysis of size distribution confirmed that size was the criterion for separation. Simultaneous DNA and RNA staining of the fractions with acridine orange and subsequent flow cytometric analysis enabled one to classify cells into resting, proliferating, and differentiating stages. Cell size was not directly correlated with proliferation in situ as determined with acridine orange flow cytometry, nor with proliferative capacity in culture as assayed by BrdU/Hoechst flow cytometry. The smallest cells, exhibiting low DNA and RNA content, which do not proliferate in vivo, required a prolonged period of serum stimulation in vitro to initiate RNA and DNA synthesis. Cells of intermediate size exhibited early RNA synthesis and maximal proliferative capacity, whereas the largest cell population displayed no RNA synthesis in culture and the least proliferative capacity. In conclusion, these results suggest that RNA synthesis early after serum stimulation, in addition to a specific, optimal cell size, correlates with the proliferative capacity of keratinocytes in cell culture.

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“…Keratinocytes were separated in a special sedimentation chamber (Tulp et al, 1980) as previously described (Pavlovitch et al, 1989). The chamber was completely filled with a dense interface liquid (fluorinated hydrocarbon; Janssen, Paris, France) at a rate of 15 ml/min.…”

Section: Low Gravity Sedimentationmentioning

confidence: 99%

“…al., 1994). Thus, opposing effects of 1,25(OH)2D3 on cell proliferation have been found, which suggests that this sterol has a complex role in the regulation of epidermogenesis.We have developed a technique which allows the isolation and quantification of the subpopulations of epidermal keratinocytes together with determination of their cycle phase by flow cytometry (Pavlovitch et al, 1989;Poot et al, 1990). In the present work such multiparameter analysis was used to study the in vivo action of vitamin D and a high calcium diet on epidermogenesis.…”

mentioning

confidence: 99%

In vivo effects of vitamin D on the proliferation and differentiation of rat keratinocytes

1996

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The hormonal form of vitamin D appears to be a physiological regulator of the epidermogenesis. While its differentiation-promoting effect is well accepted, there are conflicting reports of its action on keratinocyte proliferation. This study evaluates the specific changes induced by vitamin D treatment in the epidermis of rats nutritionally deprived of vitamin D by cell size analysis, acridine orange flowcytometry, and the immunohistochemical detection of proteins related to the different stages of differentiation (epidermal calcium binding protein and suprabasal keratins recognized by KL1 antibody) The total keratinocyte and isolated keratinocyte subpopulations were studied. Vitamin D deficiency was associated in the total population with a lower percentage of actively proliferating cells and with a lack of differentiation markers. Study of the isolated cell populations demonstrated, however, that small cells were actively proliferating, whereas they were mainly in the resting stage in the normal epidermis. Treatment with vitamin D dramatically increased cell proliferation and stimulated the appearance of differentiation markers. Some of the observed effects, such as an increase in proliferation and the appearance of epidermal calcium binding protein, were due to the normalisation of the vitamin D deficiency-induced hypocalcemia, whereas the expression of suprabasal keratins was directly dependent on vitamin D. We conclude that the action of vitamin D on the epidermis is associated with increases in both proliferation and differentiation of keratinocytes. Vitamin D itself and its resulting action on calcium homeostasis appear to contribute to the observed effects.

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Cell subpopulations within proliferative and differentiating compartments of epidermis

Cited by 13 publications

References 20 publications

Side population in adult murine epidermis exhibits phenotypic and functional characteristics of keratinocyte stem cells

Side population in adult murine epidermis exhibits phenotypic and functional characteristics of keratinocyte stem cells

Cell size and RNA content correlate with cell differentiation and proliferative capacity of rat keratinocytes

In vivo effects of vitamin D on the proliferation and differentiation of rat keratinocytes

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