摘要
通过与大鼠肝卵圆细胞的对比,对肝脏中一种新的前体细胞——小圆细胞进行分离和初步特性分析。以分步消化法制备小圆细胞,MTT法观察其增殖特点,以白蛋白、细胞角蛋白19、甲胎蛋白为标志物,应用免疫细胞化学染色的方法,对其进行鉴定。同时用DMSO刺激小圆细胞分化,检测分化结果,并与卵圆细胞加以比较。结果显示,小圆细胞增殖不明显。小圆细胞与卵圆细胞均表达白蛋白和角蛋白19。DMSO刺激后小圆细胞向卵圆细胞分化,分化前细胞AFP呈阳性,分化后细胞形状、标志物表达与卵圆细胞一致。本实验提示,小圆细胞很可能是早于卵圆细胞的一种肝脏前体细胞。
Liver undergoes profound regeneration usually after hepatic damage.It has been shown in recent study that two kinds of liver stem cells,which are mainly oval cells (OVCs) and small hepatocytes,are involved in the process of liver regeneration as they differentiated into pre- mature liver cells.However,the origination of oval cells as well as its differentiation property is not quite understood.In this study,we isolated a novel potential liver progenitor cells,namely small round cells(SRCs).The cellular features of the cells such as morphological appearance, surface maker,growth curve,and differentiation induced by DMSO were analyzed.SRCs and OVCs were obtained by using discontinuous digestions and isopyknic centrifugation,respectively. With non-radioactive measurement of MTT,the cell growth was assayed.Meanwhile,SRCs were stained with antibodies against CK19,ALB and AFP antibody to characterize their tissue-speci- ficities.The results showed that SRCs appeared in round-shaped,but irregular in size.The nuclei of SRCs were relative small with rich nucleolus visible.SRCs were semi-floated during primary culture.Even cultured in F12:DMEM(I:I) mixed-medium supplemented with 15% fetal calf serum, viability of the cells could merely be expanded over 7 days.SRCs were positively stained by CK19,ALB and AFP within initial 3 days.After DMSO stimulation,SRCs were not only morpho- logically changed into OVCs,but also expressed those markers identical to OVCs.It was suggest- ed that SRCs could be considered as an potential candidate of liver progenitors which were relat- ed to OVCs.
出处
《实验生物学报》
CSCD
北大核心
2004年第6期475-481,共7页
Acta Biologiae Experimentalis Sinica
基金
北京市重点实验室开放课题资助。
