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Optimal differentiation of in vitro keratinocytes requires multifactorial external control.

ABSTRACT: For almost 30 years, keratinocyte differentiation has been studied in numerous cell models including keratinocyte primary culture with various supplemented culture media. In this respect, it has become quite difficult to draw comparisons between studies using such a variety of culture conditions. Serum-free condition with low calcium has been used to culture basal proliferating cells, though differentiation is induced by various procedures. These latter include the addition of calcium at mM concentration and a concomitant addition of serum and calcium. Lowering the incubation temperature of cells has also been reported to induce a premature differentiation of keratinocytes in organotypic skin culture. This effect of temperature on keratinocyte differentiation has been poorly depicted, although average human skin temperature has been shown to be about 32 °C. However, studying differentiation and quantifying shifts in the differentiation rate of a cell population implies to precisely know i) the proportion of differentiated cells in the whole population, and ii) to which extent and to which level of expression, the induction of a gene or a protein might be considered as a marker of differentiation. This lack has rarely been taken into consideration and has surely led to over-interpretations of single protein induction and to consequent extrapolations to real differentiation processes. By means of paralleled analyses with immunocytofluorescence, flow cytometry, and with multiple differentiation markers quantify by qPCR and western-blot, we studied the paradoxical connection between calcium, serum, multilayer culture and incubation temperature on the differentiation of in vitro keratinocytes. Conversely to previous reports, we have shown that calcium switch is indeed a potent model for inducing calcium-dependent genes, but is not an efficient procedure when one wishes to assess the keratinocyte differentiation rate. Moreover, we have demonstrated that a synergic stimulation by calcium, serum, confluence and lower incubation temperature amplified the differentiation rate.

SUBMITTER: Borowiec AS

PROVIDER: S-EPMC3792032 | biostudies-literature | 2013

REPOSITORIES: biostudies-literature

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Optimal differentiation of in vitro keratinocytes requires multifactorial external control.

Borowiec Anne-Sophie AS Delcourt Philippe P Dewailly Etienne E Bidaux Gabriel G

PloS one 20131007 10

For almost 30 years, keratinocyte differentiation has been studied in numerous cell models including keratinocyte primary culture with various supplemented culture media. In this respect, it has become quite difficult to draw comparisons between studies using such a variety of culture conditions. Serum-free condition with low calcium has been used to culture basal proliferating cells, though differentiation is induced by various procedures. These latter include the addition of calcium at mM conc ...[more]

PMID: 24116231

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Project description:We studied previously optimal shape of external shading of windows in a cellular office with an outer edge modeled by a non-uniform rational basis spline (NURBS) curve whose control points were placed uniformly around western fin, overhang and eastern fin of the window, and whose depths were allowed to vary independently. We observed there that for each climate considered in the study there exists a shading shape close to the optimal one, but with a substantially simpler structure of control points for the NURBS curve. This simpler structure was reflected in partitioning control points into six groups such that all control points in the same group have equal depths, with groups corresponding to lower part of the western fin, upper part of the western fin, joint of the western fin and the overhang, internal part of the overhang, joint of the overhang and the eastern fin and the remaining part of the eastern fin. Here we confirm that shadings with control point structure restricted in such way can perform as well as shadings with unrestricted control points by optimising shape of external shading of windows in an apartment room for both restricted and unrestricted control point structure for the same range of climates, and showing that differences in heating and cooling demands between Pareto optimal shadings in both cases are negligibly small. This grouping of control points thus gives a simple and natural division of shading into a small number of basic constituents that have most impact on its heating and cooling demands. We further consider the convex hull of the Pareto front for shadings with restricted control points, as it contains shadings that minimise equivalent source energy in terms of the ratio of efficiencies and source energy conversion factors for district heating and cooling. We show that, in cases when depths of control point groups in convex hull shadings do not experience sudden changes between their extremal values, these depths can be fitted reasonably well by a sigmoid function that results in functional shadings that satisfactorily approximate heating and cooling demands of shadings in the Pareto front.

Dataset Information

Optimal differentiation of in vitro keratinocytes requires multifactorial external control.

Publications

Optimal differentiation of in vitro keratinocytes requires multifactorial external control.

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