Hepatic precursors derived from murine embryonic stem cells contribute to regeneration of injured liver
Hepatic precursors derived from murine embryonic stem cells contribute to regeneration of injured liver
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DOI：
10.1002/hep.21441
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发表时间：
2006-12-01
期刊：
影响因子：
13.5
通讯作者：
Thorgeirsson, Snorri S.
中科院分区：
文献类型：
--
作者：
Heo, Jeonghoon;Factor, Valentina M.;Thorgeirsson, Snorri S.
我们建立了一个有效的系统，用于从小鼠胚胎干细胞（ES）中分化、扩增和分离肝祖细胞，并评估其重建受损肝脏的能力。利用白蛋白（ALB）增强子/启动子调控的绿色荧光蛋白（GFP）报告基因转染的小鼠ES细胞，我们发现无血清化学成分确定的培养基支持胚状体（EBs）的形成和肝系细胞在缺乏外源生长因子或饲养细胞层的情况下的分化。在EB培养7天后，在接近“跳动”肌细胞的剂量下检测到第一个表达ALB的GFP（+）细胞。GFP+细胞数量增加，获得肝细胞样形态和肝细胞特异性标志物（即，ALB、AAT、TO和G6 P），并且到28天代表了从EB副产物分离的超过30%的细胞。流式细胞术纯化的GFP（+）细胞移植到MUP-uPA/SCID小鼠体内后，可发育成功能性肝细胞，无细胞损伤，并参与了病变肝脏的修复。ES细胞衍生的肝细胞对正常的生长调节有反应，并且在CCl 4诱导的肝损伤的额外生长刺激后以与宿主肝细胞相同的速率增殖。移植的GFP+细胞也分化为胆管上皮细胞。总之，一个高度富集的人口的定向肝细胞前体细胞可以产生从ES细胞在体外有效的细胞替代疗法。
We established an efficient system for differentiation, expansion and isolation of hepatic progenitor cells from mouse embryonic stem (ES) cells and evaluated their capacity to repopulate injured liver. Using mouse ES cells transfected with the green fluorescent protein (GFP) reporter gene regulated by albumin (ALB) enhancer/promoter, we found that a serum-free chemically defined medium supports formation of embryoid bodies (EBs) and differentiation of hepatic lineage cells in the absence of exogenous growth factors or feeder cell layers. The first GFP(+) cells expressing ALB were detected in dose proximity to "beating" myocytes after 7 days of EB cultures. GFP+ cells increased in number, acquired hepatocyte-like morphology and hepatocyte-specific markers (i.e., ALB, AAT, TO, and G6P), and by 28 days represented more than 30% of cells isolated from EB outgrowths. The FACS-purified GFP(+) cells developed into functional hepatocytes without evidence of cell ftision and participated in the repairing of diseased liver when transplanted into MUP-uPA/SCID mice. The ES cell-derived hepatocytes were responsive to normal growth regulation and proliferated at the same rate as the host hepatocytes after an additional growth stimulus from CCl4-induced liver injury. The transplanted GFP+ cells also differentiated into biliary epithelial cells. In conclusion, a highly enriched population of committed hepatocyte precursors can be generated from ES cells in vitro for effective cell replacement therapy.