Mesangioblast clone comparison- U74C

Mouse dorsal aorta from E9.5 mouse embryos (C57BL6) was grown as an explant culture as described elsewhere (De Angelis et al., 1999). After 5 days the explant was dissociated into a single cell suspension, and plated at limiting dilution on a feeder layer of Mitomycin C-treated primary embryonic mouse fibroblasts, in 96 multiwell plates in complete medium, RPMI 20% Fetal Calf Serum. After 1 week, mesangioblast clones appeared approximately 2-4% of the wells. When the clones had grown to approximately 103 cells they were passed on gelatin-coated dishes, and further expanded. Clones D16 and D351 were used for microarray analysis. Keywords: other

取C57BL/6品系小鼠胚胎发育第9.5天的背主动脉（mouse dorsal aorta），参照已发表方法（De Angelis等，1999）进行外植体培养。培养5天后，将外植体消化为单细胞悬液，以有限稀释法接种于丝裂霉素C（Mitomycin C）处理的原代小鼠胚胎成纤维细胞滋养层上，使用含20%胎牛血清(Fetal Calf Serum)的RPMI完全培养基，在96孔板中培养。培养1周后，约2%~4%的孔中出现系膜祖细胞（mesangioblast）克隆。当克隆生长至约1000个细胞时，将其传代至明胶包被培养皿中并进行进一步扩增。选取克隆D16与D351进行基因芯片（microarray）分析。关键词：其他