【摘要】 用 型胶原酶消化和微孔过滤法分离新生大鼠肺微血管段的内皮细胞和周细胞 ,再用富含 PDGF的选择性条件培养液刺激周细胞的生长和抑制内皮细胞生长而建立了肺血管壁周细胞的培养方法。 S180细胞条件培养液(SCCM)的灭活 ,分离、纯化微血管段 ,静置及筛选周细胞是获得新生大鼠肺血管壁周细胞纯培养的关键。用该方法培养的第 5代周细胞 ,经相差显微镜和免疫细胞化学法检查 ,纯度达 98%以上 ;细胞倍增时间为 5 4.3h,且在 12代以内 ,细胞形态结构特征保持稳定 ,适合做周细胞的体外实验研究更多还原

【Abstract】 Endothelial cells and pericytes in microvascular segments were dissociated by type Ⅳ collagenase digestion and filtering through screens , then selective conditioned media rich in platelet derive growth factor(PDGF) was used to stimulate the growth of pericytes and to inhibit the growth of endothelial cells. The culture method for pulmonary microvascular pericytes in newborn rats was established.The key points for obtaining pure culture of pericytes were as follows: 1)inactivation of conditioned medium from mouse sarcoma 180 cells (SCCM); 2) separating and purifying the microvascular segments; 3) keeping the culture dishes of the primary culture still in the first few days; and 4) screening pericytes etc. Phase-contrast microscopic examination and immunocytochemical staining tests showed that the purity of cultured pericytes at the 5th passage from this method was over 98 %; The doubling time for the cultured pericytes was 54.3 h, and morphological charateristics of the cells at 5th to 12th passages were stable. The cultured pericytes from this method were applicable to experimental investigation in vitro .更多还原