一种改良成人肺动脉平滑肌细胞的培养方法

An improved method of human pulmonary smooth muscle cells culturein vitro

目的:探讨成人肺动脉平滑肌细胞(pulmonary arterial smooth muscle cells,PASMCs)体外培养及培养效率的方法。方法:在无菌条件下,取3cm左右肺动脉,去除内膜后,剪成1mm×1mm大小的组织块,贴块法培养,在高糖DMEM/F12 1∶1培养基中加入血小板衍生长因子(PDGF)10μg/L培养hPASMCs,采用倒置显微镜观察细胞形态,免疫荧光法鉴定α-平滑肌肌动蛋白(smooth muscle-α-actin,SM-α-actin)和钙调蛋白calponin。结果:采用改良组织块培养,克服了hPASMCs体外培养休眠期过长的缺点,稳定获得纯度达98%以上,细胞呈长梭型,生长致密时排列成相互平行的束状,未见明显异形的细胞,SM-α-actin和calponin阳性。结论:该改良方法能成功分离和培养出纯度较高的成人肺动脉平滑肌细胞,且重复性好,细胞常量大,简便而有效。

Objective： To study the method of human pulmonary smooth muscle cells （hPASMCs） culture in vitro Methods： A sample of 3 cm long sap pulmonary artery was harvested under sterile condi- tions. After the internal and external membrane were removed, the middle membranes were cut into 1 mm X 1 mm size pieces, and were cultured by tissue-piece inoculation method, added to DMEM medium with PDGF （10 tg/L） and high concentration of glucose （DMEM/F12 1 ： 1）. The smooth muscle cells were identified by immunohistochemistry. Results： Culturing with PDGF and high glucose in vitro over- comes the short coming of the long period of hPASMC dormancy, and the purity of the third passage hPASMCs was more than 98. The cultured cells owed the typical long spindle, peak and valley appear-