Gene expression profile of macrophages of monocytic origin and Kupffer cells under conditions of co-cultivation with Ito cells, endotheliocytes of sinusoidal capillaries of the liver and HGF.
The aim of the work was to study the expression profile of Kupffer cells and macrophages of monocytic origin under conditions of co-cultivation with Ito cells, endotheliocytes of sinusoidal capillaries of the liver and HGF. To test the hypothesis that it is the microenvironment that forms the specific phenotype of resident organ macrophages. We used microarrays for comparison of Kupffer cells and macrophages of monocytic origin gene expression on different time periods of cocultivation with Ito cells, endotheliocytes of sinusoidal capillaries of the liver and HGF.
Overall design
Kupffer cells were obtained by magnetic sorting from the liver of Balbc male mice by the F4/80 antigen, and monocytes from the blood of the same mice by the CD115 marker. Next, the cells were preliminarily cultured for a 1 day with MCSF. Next, monocytic macrophages and Kupffer cells were co-cultivated for 1 or 3 days with Ito cells or sinusoidal endotheliocytes or HGF. Macrophages of monocytic origin or Kupffer cells that continued to be cultivated for 1 or 3 days in a medium containing MCSF served as controls. At the end of the required period, RNA was extracted from the cells, which was used for transcriptome analysis.
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