摘要
小鼠骨髓内皮细胞系建立、保存和传代培养已10年之久。本研究的目的是对该内皮细胞系的生物学特性进行新的鉴定,以便利用该内皮细胞系进行进一步研究。取经传代培养的该内皮细胞系细胞,进行显微光镜和电子显微镜观察、分子标志检测、吞噬实验、生长动力学分析以及核型鉴定等。结果表明:细胞在亚汇合贴壁层中多为椭圆形、圆形或梭形,部分可呈条索状排列,在汇合层中一般为鹅卵石状。CD31、vWF抗体染色阳性细胞占94%以上,摄取DiI-Ac-LDL的细胞达98.5%。在常规培养条件下,快速增殖期的细胞群体倍增时间约为54.6小时,分裂指数最高达到38‰。集落产率介于4.33%-7.40%。核型主要为超二倍体。结论:该细胞系保持了内皮细胞的基本特征,生长动力学和一些生物学行为与建系之初稍有差异。
The murine bone marrow endothelial cell line （mBMEC） has been maintained by means of subculture and cryopreservation for over 10 years since it was established in our laboratory. This study was aimed to newly identify biological characteristics of this cell line for further study. The cultured mBMEC cells were observed by inverted microscopy and transmission electron microscopy （TEM）. PECAM-1 （CD31） and von Willebrand factor （vWF） were detected by immunofluorescent staining. The phagocytotic activity of the cells in culture was tested by using fluorescent acetylated low-density lipoprotein （Dil-Ac-LDL）. The cell growth kinetics analysis and karyotype analysis were performed. The results showed that the adherent cells were mostly elliptical, rounded and spindle-shaped, and some of them connected to each other to form cord- and network-like arrangements in mBMEC cultures at subconfluence. The adherent cells grew up to confluence as a cobblestone-like monolayer. Several ultrastructural features of the endothelial cells could be observed in TEM sections of the cultured cells. More than 94% of mBMEC cells were positive for either CD31 or vWF. The phagocytotic ingestion of Dil-Ac-LDL occurred in 98.5% of cells. In normal culture conditions, the cells grew with a mean population doubling time of 54.6 hours and the maximal mitotic index was 38%0 in the rapid growth period. The colony yields were 4.33% to 7.40% depending on the plating density of cells. Karyotypes of all the cells were aneuploidy with a greater percentage of hyperdiploid. It is concluded that mBMEC cells retain the fundamental properties of endothelial cells, but the growth kinetics and biological behaviors are slightly different from those in the early days after the establishment of this cell line.
出处
《中国实验血液学杂志》
CAS
CSCD
2007年第6期1231-1235,共5页
Journal of Experimental Hematology
基金
国家自然科学基金面上项目资助
编号30470637
关键词
内皮细胞系
传代培养
生物学特性
endothelial cell fine
subculture
biological characteristics
