Ionocytes were obtained from the FGF10CreERT2:Rosa26tdTomato and MECs from the αSMA-GFP mice by FACS as described above (Zyrianova et al., 2019 (link)). Purified ionocytes were plated into the center of each well of a four-well chamber slide coated with a thin layer of Matrigel. When cells attached, a 10 μL of low growth factors Matrigel drop was gently spreaded as a thin layer on a top of attached and placed into cell incubator at 37C for 5–10 min. When gel settled, purified MECs have been plated on a top of Matrigel and cells were grown in the defined medium: high glucose DMEM (Thermo Fisher Scientific, # 11995073) supplemented with 0.1% Lipid-Rich Bovine Serum Albumin (AlbuMAX I, Thermo Fisher Scientific, ##11020), insulin-transferrin-selenium (Thermo Fisher Scientific, #41400-045), human transferrin (40 μg/mL, Thermo Fisher, #0030124SA) and freshly prepared ascorbic acid (150 μg/mL). MEC cultures on the Matrigel without ionocyte were used as controls. Cells were grown for 48 h and stained with the phosphor-histone-3 antibody and DAPI to visualize proliferating cells.
Lipid rich bovine serum albumin albumax 1
Manufactured by Thermo Fisher Scientific
Lipid-Rich Bovine Serum Albumin (AlbuMAX I) is a cell culture supplement derived from bovine serum. It is a protein-rich solution that contains a high concentration of lipids.
Automatically generated - may contain errors
2 protocols using lipid rich bovine serum albumin albumax 1
1
Ionocyte-MEC co-culture protocol
2
Ionocyte-MEC co-culture protocol
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Ionocytes were obtained from the FGF10CreERT2:Rosa26tdTomato and MECs from the αSMA-GFP mice by FACS as described above (Zyrianova et al., 2019 (link)). Purified ionocytes were plated into the center of each well of a four-well chamber slide coated with a thin layer of Matrigel. When cells attached, a 10 μL of low growth factors Matrigel drop was gently spreaded as a thin layer on a top of attached and placed into cell incubator at 37C for 5–10 min. When gel settled, purified MECs have been plated on a top of Matrigel and cells were grown in the defined medium: high glucose DMEM (Thermo Fisher Scientific, # 11995073) supplemented with 0.1% Lipid-Rich Bovine Serum Albumin (AlbuMAX I, Thermo Fisher Scientific, ##11020), insulin-transferrin-selenium (Thermo Fisher Scientific, #41400-045), human transferrin (40 μg/mL, Thermo Fisher, #0030124SA) and freshly prepared ascorbic acid (150 μg/mL). MEC cultures on the Matrigel without ionocyte were used as controls. Cells were grown for 48 h and stained with the phosphor-histone-3 antibody and DAPI to visualize proliferating cells.
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