兔骨髓单个核细胞体外向内皮细胞诱导的实验研究Research of endothelial cells induced from rabbit marrow mononuelear cells in vitro
谭洪波;杨柳;段小军;张洪鑫;左镇华;金旭红;李忠;王富友;
摘要(Abstract)：
[目的]研究兔骨髓单个核细胞(marrowmononuclearcells,MNCs)向内皮方向诱导的简捷有效方法,进行科学的计量分析并观察体外传代培养条件下的主要生物学特点。[方法]梯度离心分离兔MNCs,直接诱导培养,以每106个MNCs收获第1、2代内皮细胞的数量计量内皮细胞的收获效率;传代培养,倒置显微镜观察细胞形态及生长特点,免疫组化及内皮细胞功能鉴定。[结果]以1×106/cm2的密度接种MNCs,经7~8d培养可收获第1代内皮细胞;第1、2代内皮细胞的收获总量和MNCs的收获量呈显著的正相关,每106个MNCs平均可收获第1代内皮细胞约(7·086±0·75)×104个,平均收获第2代细胞(53·20±6·47)×104个。诱导的内皮细胞为铺路石样,呈单层生长,第2代细胞CD31、八因子相关抗原免疫组化均为阳性,摄取低密度脂蛋白和结合凝集素实验均阳性。[结论]以1×106/cm2的MNCs接种密度,收获第1、2代内皮细胞所需时间较短,收获效率高,在体外培养条件下生物学特性稳定,有望作为骨组织工程血管化的种子细胞。
关键词(KeyWords)： 内皮细胞;组织工程;细胞培养;种子细胞
基金项目(Foundation): 国家自然科学基金资助项目(30300079)
作者(Author): 谭洪波;杨柳;段小军;张洪鑫;左镇华;金旭红;李忠;王富友;
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