Prospective isolation of a bipotential clonogenic liver progenitor cell in adult mice
Prospective isolation of a bipotential clonogenic liver progenitor cell in adult mice
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DOI：
10.1101/gad.2029411
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发表时间：
2011-06-01
影响因子：
10.5
通讯作者：
Grompe, Markus
中科院分区：
文献类型：
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作者：
Dorrell, Craig;Erker, Laura;Grompe, Markus
由于缺乏真正特异的标记物，成人肝干/祖细胞的分子鉴定一直受到阻碍。为了分离假定的成年肝祖细胞，我们使用细胞表面标记抗体，包括MIC1-1C3，从正常成年小鼠或经历卵圆细胞反应的小鼠中分离出肝细胞亚群，并测试它们在体外形成双系集落的能力。在正常或卵圆细胞损伤的肝脏中，强大的克隆形成活性被发现仅限于抗原性定义为CD45(-)/CD11b(-)/CD31(-)/MIC1-1C3(+)/CD133(+)/CD26(-)，的胆管细胞亚群，频率分别为34或25之一。基因表达分析表明，Sox9仅在正常肝细胞的这一亚群中表达，并且相对于受损肝脏中的其他细胞组分，Sox9高度浓缩。使用Sox9creer(T2)-R26R(YFP)小鼠的活体谱系追踪显示，在祖细胞驱动的肝再生过程中增殖的细胞是表达Sox9的前体细胞的后代。对正常和DDC(3，5-二乙氧基-1，4-二氢碰撞碱或diethyl1，4-dihydro-2，4，6-trimethyl-3，5-pyridinedicarboxylate)-treated)富集祖细胞和去祖细胞的基因表达进行了全面的阵列比较，发现了新的潜在的肝祖细胞调控因子。
The molecular identification of adult hepatic stem/progenitor cells has been hampered by the lack of truly specific markers. To isolate putative adult liver progenitor cells, we used cell surface-marking antibodies, including MIC1-1C3, to isolate subpopulations of liver cells from normal adult mice or those undergoing an oval cell response and tested their capacity to form bilineage colonies in vitro. Robust clonogenic activity was found to be restricted to a subset of biliary duct cells antigenically defined as CD45(-)/CD11b(-)/CD31(-)/MIC1-1C3(+)/CD133(+)/CD26(-), at a frequency of one of 34 or one of 25 in normal or oval cell injury livers, respectively. Gene expression analyses revealed that Sox9 was expressed exclusively in this subpopulation of normal liver cells and was highly enriched relative to other cell fractions in injured livers. In vivo lineage tracing using Sox9creER(T2)-R26R(YFP) mice revealed that the cells that proliferate during progenitor-driven liver regeneration are progeny of Sox9-expressing precursors. A comprehensive array-based comparison of gene expression in progenitor-enriched and progenitor-depleted cells from both normal and DDC (3,5-diethoxycarbonyl-1,4-dihydrocollidine or diethyl1,4-dihydro-2,4,6-trimethyl-3,5-pyridinedicarboxylate)-treated livers revealed new potential regulators of liver progenitors.