摘要
培养表达人apoAI转基因小鼠主动脉平滑肌细胞(SMC)。从具有可诱导,高效,多组织表达人apoAI转基因小鼠分离主动脉,用改良贴块法培养血管SMC。细胞经相差显微镜和免疫细胞化学证实具有SMC特征。Northernblot表明转基因鼠主动脉平滑肌细胞中有人apoAImRNA表达,而正常鼠则未检测出,为在体外从细胞和分子水平研究anOAI基因表达在胆固醇代谢及动脉粥样硬化发生中的作用提供了较理想的细胞模型。
Culture of smoothe muscl cells originally from the aorta (VSMCs) human apolipoprotein Al (apo Al ) transgenic mice was established. Fresh heaft and aortac specimens were perfused immediately after the removal with trypsin at low concentration. The aorta were then cut into small pieces: washed with culture medium; and cultured with DMEM containing 20% fetal calf serum with 100U/mlpenicillin and 100ug/mL streptomycin. Under phase contrast microscope the cultured cells possessed, peak and valley characteristics for smooth muscle cells.Immunocytochemical staining with mAb against mouse smooth muscle a-actin demonstrated these cells were possitive. Noythern blot analysis using human apoAI gene as the probe showed expression of human apoAI mRNA in the SMC from transgenic mice, but undetectable in the ASMCs of the control mice. It provides an ideal cell model in studying the role of apo Al in lipid metabolism and atherogenesis.
出处
《基础医学与临床》
CSCD
2000年第1期38-40,共3页
Basic and Clinical Medicine
关键词
载脂蛋白AI
转基因小鼠
平滑肌细胞
细胞培养
apolipoprotcin AI
the cultured treansgenic mice
smooth muscle cells
cell culture