摘要
目的体外培养小鼠胎肝细胞，探索其生长和分化模式，并进行干细胞特征的表型鉴定。方法分离培养不同时期小鼠胎肝细胞，动态观察细胞形成的集落数及生长模式；收获集落进行干细胞表面标记物流式检测；传代培养胎肝细胞，探索甲胎蛋白（AFP）变化趋势及细胞分化模式。结果经体外培养，第15天胎肝细胞形成明显集落，生长状况最好，干细胞的表面标记提示形成的集落具有肝干细胞特性。传代培养中，第15天胎肝干细胞以线性模式生长；分化模式中AFP呈现正态分布变化。结论随胎龄增加，有干细胞特性的细胞逐渐减少。体外培养中，肝干细胞呈线性模式增殖，逆向线性方式分化。
Objective To study the proliferation and differentiation patterns of hepatic stem cells from mice cultured in vitro isolated identify their characteristics. Methods Mice were divided in to 5 groups according to the pregnant （embryo days, ED13, ED15 and ED18） or born age （day0 and 3）. Hepatic stem cells were isolated and cultured in vitro. The amount of the stem cells as well as their growing situation and differentiation pattern were observed and compared among different groups and markers of stern cells （CD117, CDg0.1, CD49f, c -Met）, hepatic cells（AFP） were used to identify the cultures. Results The cells with best situation of growing as colonies were obtained from ED15 group. Their expression of specific markers suggested that they were hepatic stem cells. The stem cells isolated from ED15 mice in subcuhuring proliferated in line pattern and differentiated in reverse line pattern. The expression of AFP varied as normal distribution as cell differentiation development. Conclusion Most cells have characteristics of hepatic stem cells isolated from fetal liver and the number of these cells decreases gradually as embryo duration prolongs. The hepatic stem ceils proliferate in line pattern and differentiate into hepatic cells after in vitro culture.
出处
《中华肝胆外科杂志》
CAS
CSCD
北大核心
2009年第10期768-771,共4页
Chinese Journal of Hepatobiliary Surgery
基金
国家自然科学基金（编号：30772102）
关键词
肝干细胞
生长
分化
鉴定
Hepatic stem cell
Proliferation
Differentiation
Identification
