摘要
目的:以鼠尾胶为贴黏剂,建立一种成本低且生长状态良好的微血管内皮细胞体外培养体系。方法:无菌条件下制作鼠尾胶,并均匀铺在培养皿中。分离收集脑微血管段,接种于铺有鼠尾胶的培养皿中进行培养。用内皮细胞的两种重要标记物VIII因子相关抗原和γ-谷氨酰转酞酶(-γglutamyl-transpeptidase,γ-GT)鉴定细胞,并以3H-TdR掺入法观察培养细胞对血清刺激的增殖反应性。结果:显微镜观察从脑微血管段生长出的细胞具有内皮细胞生长特性:单层“卵石样”排列特征和接触生长抑制;VIII因子免疫组织化学染色呈阳性;培养上清含有大量的γ-谷氨酰转酞酶[(5.01±0.07)IU/L];证实鼠尾胶做为贴黏剂获得的是内皮细胞。10%胎牛血清刺激后3H-TdR掺入率较对照组增加87.5%(P<0.05)。结论:本研究用鼠尾胶做为贴黏剂,降低微血管内皮细胞培养成本,且细胞生长状态良好,表明该方法是一种比较经济可靠的微血管内皮细胞培养体系。
Objective： To established a low cost in vitro culture model of microvascular endothelial cells （MEC） by using rat tail collagen as an adhesive reagent. Methods： Rat tail collagen was produced under the sterile condition. The culture dishes were covered with rat tail collagen. Microvessles were isolated, collected and cultured in the culture dishes. MEC were identified using two markers, including factor VIII antigen and γ-glutamyl-transpeptidase（γ-GT）. In addition, the responsiveness of MEC to fetal serum was observed by measuring ^3H thymidine incorporation. Resuits： MEC grew out as typical ＂cobblestone＂ and contact inhibition under an inverted microscope. The cells were positive for factor Ⅷ antigen. And a lot of γ-GT was secreted in the supernatants of the cultured cells. Additionally, ^3H thymidine incorporation of the ceils treated by 10% fetal serum elevated by 87.5% （vs control, P〈0.05）. Conclusions： Using rat tail collagen as an adhesive reagent, the cost of MEC culture was decreased and MEC had a better growing state. These results indicated that the method is an economic and credible cultural system of MEC.
出处
《军医进修学院学报》
CAS
北大核心
2005年第6期464-465,共2页
Academic Journal of Pla Postgraduate Medical School
基金
国家自然科学基金(30370569)
解放军总医院科研基金(01YM30)
关键词
鼠尾胶
黏着剂
毛细血管
细胞培养
rat tail collagen
adhesives
capillaries
cell culture
