Kubrycht J, Maxová H, Nyč O, Vajner L, Novotná J, Hezinová A, Trnková A, Vrablová K, Vytášek R, Valoušková V. Physiol Res. 2010 Jun 9. IF: 1.653
Abstract:
Prolonged cultivation of separated rat lung mast cells (LMC) in vitro is necessary to better investigate a possible role of LMC in different time of model tissue remodeling induced by hypoxia. Consequently, rat lung mast cells (LMC) were separated here using protocol including an improved proteolytic extraction and two subsequent density gradient separations on Ficoll-Paque PLUS and a new generation of Percoll, i.e. Percoll PLUS. Instead of usual isotonic stock Percoll solution, an alternative asymptotically isotonic stock solution was more successful in our density separation of LMC on Percoll PLUS. Separated cells were cultivated six days in media including stem cell factor, interleukins 3 and 6, and one of two alternative mixtures of antibiotics. These cultivations occurred without any contamination and with only low frequent changes in cell size and morphology. Model co-cultivation of two allogenic fractions of LMC caused frequent and sometimes rapid and considerable changes in cell morphology and size. In contrast to these observations no or rare morphological changes were found after cultivation under hypoxic conditions.
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