%0 Journal Article %T Sedimentation field-flow fractionation separation of proliferative and differentiated subpopulations during Ca2+-induced differentiation in HaCaT cells. %+ Biomolécules Thérapies anti-tumorales (EA4021) %+ Homéostasie Cellulaire et Pathologies (HCP) %+ Université de Limoges (UNILIM) %A Micallef, Ludovic %A Battu, Serge %A Pinon, Aline %A Cook-Moreau, Jeanne %A Cardot, Philippe %A Delage, Christiane %A Simon, Alain %< avec comité de lecture %Z ACL-51 %@ 1570-0232 %J Journal of Chromatography B - Analytical Technologies in the Biomedical and Life Sciences %I Elsevier %V 878 %N 15-16 %P 1051-8 %8 2010-05-01 %D 2010 %R 10.1016/j.jchromb.2010.03.009 %M 20371217 %Z Life Sciences [q-bio]/Cellular Biology %Z Life Sciences [q-bio]/Pharmaceutical sciencesJournal articles %X The spontaneously immortalized human keratinocyte cell line HaCaT is widely used as a human keratinocyte model. In a previous comparative study between normal human keratinocytes (NHKs) and HaCaT, we reported that Ca2+ concentrations greater than 1mM induced differentiation in vitro in both cell types, notably characterized by increased expression of differentiation markers keratins 1 (K1), 10 (K10) and involucrin. Surprisingly, cells had a higher proliferative activity than those cultured with low Ca2+ levels. These results raised many questions; in particular concerning the emergence of HaCaT cells subpopulation which would have different differentiation states and/or proliferation rates throughout Ca2+-induced differentiation. To isolate these subpopulations, we used sedimentation field-flow fractionation (SdFFF). Results demonstrated that the most differentiated cells (HC-F1), characterized by the highest expression of keratinocyte differentiation markers, had the lowest proliferative activity. In contrast, less differentiated cells (HC-F2) maintained a higher proliferative activity. SdFFF is a tool to sort differentiated and/or proliferating cells from a total pool previously treated with a Ca2+ concentration inducing differentiation, and can be use to prepare biological models necessary for studying HaCaT cell proliferation after Ca2+-induced differentiation treatment. %G English %L hal-00597638 %U https://unilim.hal.science/hal-00597638 %~ UNILIM %~ CNRS %~ EA3842 %~ LCSN %~ GEIST %~ CAPTUR %~ PEIRENE %~ OMEGAHEALTH