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目的探讨大鼠肺微血管内皮细胞培养方法 ,并期望发展一种简便、实用的体外培养方法。方法用 7%O2 (4h/d ,3d)诱导培养猪肺动脉内皮细胞 ,然后收集该低氧诱导的培养基后加入大鼠肺边缘组织块 ,再培养微血管内皮细胞。结果加入低氧诱导培养基后 48h ,即有明显细胞增生 ;原代培养 1wk后 ,细胞呈典型的鹅卵石状连片生长 ,继续可以传代 3～ 4次。与对照方法相比 ,采用低氧诱导培养基法培养大鼠肺微血管内皮细胞 ,贴块后细胞增生反应较早出现、繁殖较快 ,细胞老化延迟 ,传代后细胞生长良好 ,3代以内细胞形态无明显退变。结论本文探讨的低氧诱导培养基方法 ,具有简便、实用的特点 ,可用于建立大鼠肺微血管内皮细胞的细胞模型 ,用于高原医学研究。
Objective To explore the method of culture of rat pulmonary microvascular endothelial cells and to develop a simple and practical method of in vitro culture. Methods Porcine pulmonary artery endothelial cells were induced by 7% O2 (4h / d, 3d). The hypoxia-inducible medium was collected and then added to the marginal tissue of rat lungs for further culture of endothelial cells. Results 48h after adding hypoxia induction medium, obvious cell proliferation; primary culture 1wk, the cells showed a typical pebble-like contiguous growth, can continue to pass 3 to 4 times. Compared with the control method, the rat pulmonary microvascular endothelial cells were cultured by hypoxia-induced medium, the cell proliferation reaction appeared earlier, the propagation was rapid, the cell aging was delayed, the cell growth was good after passage, the cell morphology within 3 generations No obvious degeneration. Conclusion The hypoxia-inducible medium method discussed in this paper is simple and practical. It can be used to establish a cell model of rat pulmonary microvascular endothelial cells for plateau medical research.