比较两种内皮祖细胞增殖、抗凋亡和集落形成能力  

The Differences of Proliferative Potential,Apoptosis Resistance and Colony Forming Capacity between Two Types of Endothelial Progenitor Cells

作　　者：段华新[1] 卢光琇[2] 程腊梅[2] 

机构地区：[1]广州医学院附属市一人民医院血液科,广东广州510180 [2]中南大学生殖与干细胞工程研究所人类干细胞国家工程研究中心,湖南长沙410078

出　　处：《广州医学院学报》2007年第4期57-61,共5页Academic Journal of Guangzhou Medical College

基　　金：国家863项目基金(2003AA205181);国家973项目基金(00CB51010);国家自然科学基金重点项目(30030070)

摘　　要：比较两种内皮祖细胞的增殖潜能、抗凋亡和集落形成能力。方法：本研究用增殖曲线、倍增水平、倍增时间的计算及MTT增殖实验等方法比较两种内皮祖细胞的增殖特点;用单个细胞的集落形成实验比较两种细胞的集落形成能力;并用流式细胞术检测两种细胞凋亡发生率。结果：循环成血管细胞（CACs）只能达到4-6个倍增,而高增殖潜能内皮祖细胞（HPP-EPCs）能达到60多个倍增。HPP-EPCs的累计倍增水平是CACs的9.53倍。MTT实验的结果也支持HPP-EPCs增殖明显快于CACs。来自CACs的单个细胞不能形成集落。但是,HPP-EPCs的单个细胞中45%在14 d的培养期间形成了集落,并且来自二级集落的96个单个细胞中有5个三级集落形成。HPP-EPCs显示比CACs具有更强的抗凋亡能力。结论：HPP-EPCs比CACs具有更强的增殖能力、集落形成能力和抗凋亡能力。Objective：To compare the proliferative characteristics between the two types of endothelial progenitor cells （EPCs）. Methods： In order to compare their growth characteristics, the growth curves were generated and the population doubling time and cumulative population doubling levels were calculated for the two types of cells. Flow cytometry was performed to analyze the apoptosis in two types of EPCs induced by 48 h serum-free culture. Results： Circulating angiogenic cells （ CACs ） could only reach 4 - 6 population doublings but high proliferative potentialendothelial progenitor cells （HPP-EPCs） could reach over 60 population doublings without any sign of senescence in vitro. The average cumulative population doubling levels of HPP-EPCs was 9.53-fold higher and the average population doubling time was shorter than those of CACs. The result of MTT assay showed that the optical density A490nm values of HPP-EPCs were about 4-fold higher than those of CACs after 24 h culture with the same number of seeded cells. The experiments of colony forming showed that no colony was formed in 960 individual cells from CACs, whereas more than 35 percent of 960 individual cells from HPP-EPCs colonies survived and differentiated into 2-50 progeny cells, and about 45 percent of 960 individual cells formed colonies with more than 50 cells in a 2-wk culture period. And 5 tertiary colonies were formed in the total 96 individual cells from the secondary colonies. These data suggested that we have cultured different hierarchies of EPCs from human umbilical cord blood including CACs and HPP-EPCs, and the later contained high proliferative potential-endothelial colony forming cells and low proliferative potential-endothelial colony forming cells. The results of flow cytometry showed that 1.2% ±0.6% early-stage apoptosis cells and 0.1% ± 0.1% late-stage apoptosis cells were detected in HPP-EPCs after 48-h serum-free induction whereas 4.5% ± 1.4% early-stage apoptosis and 2. 1% ± 0. 8% late-stage apoptosis in CACs. Concl

关 键 词：内皮祖细胞 增殖能力 凋亡 集落形成 

分 类 号：Q253[生物学—细胞生物学]