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.
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Serum-free condition with low calcium has been used to culture basal proliferating cells, though differentiation is induced by Norway gentlemen club Mandal 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.
However, studying differentiation and quantifying shifts in the differentiation rate of a cell population implies to precisely know i the proportion Average breast size in Kristiansund 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 Watermelon girl Bodo 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. This is an open-access article distributed under the terms Elegant asian Horten the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
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The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. Competing interests: The authors have declared that no competing interests exist. Epidermis is the outer compartment of skin and is formed by stratified layers of keratinocytes from the basal proliferating compartment up to the spinous, the granular and the Kristinasund layers.
For almost 30 years, keratinocyte differentiation has been studied with numerous cell models including orthotypic culture [ 12 ] and primary Krisstiansund culture.
In order to simplify culture procedures, the latter has been developed by removing the fibroblast-based feeder layer of a stratified culture. The mono-layered primary culture of keratinocytes has also let researchers to experiment on living cells.
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In this respect, it has become quite difficult to make comparisons between the studies using such a variety of culture conditions. It has thus become easy to get lost in this jungle of procedures and to formulate wrong interpretations.
Strikingly, an original article demonstrated that optimal rate of keratinocyte proliferation was reached at about 0. The induction of keratinocyte differentiation is even more confusing. Experiment was performed three times independently.
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