Purification of fetal liver stem/progenitor cells containing all the repopulation potential for normal adult rat liver
Purification of fetal liver stem/progenitor cells containing all the repopulation potential for normal adult rat liver
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DOI：
10.1053/j.gastro.2008.01.007
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发表时间：
2008-03-01
期刊：
影响因子：
29.4
通讯作者：
Shafritz, David A.
中科院分区：
文献类型：
--
作者：
Oertel, Michael;Menthena, Anuradha;Shafritz, David A.
背景和目标：在此之前，我们发现移植未分离的胚胎日（艾德）14胎肝干/祖细胞（FLSPC）后，可以实现高水平、长期的肝脏替代。然而，对于临床应用，移植高度富集的细胞，同时保持高的再增殖潜力将是至关重要的。方法：Dlk-1是细胞表面跨膜蛋白δ样家族的成员，在人和啮齿动物胎肝中高度表达。用免疫磁珠法从ED_（14）胎肝中分离Dlk-1（+）细胞，体外检测其肝基因表达谱和特性，以及移植到正常成年大鼠肝中后的体内增殖和分化潜能。结果如下：将大鼠ED 14 FLSPC纯化至95%的均一性，并表现出预期的肝干/祖细胞的细胞培养和基因表达特征。大鼠ED 14 FLSPC为甲胎蛋白（+）/细胞角蛋白-19（+）或甲胎蛋白（+）/细胞角蛋白-19（-），并含有胎肝中发现的所有正常肝脏再增殖能力。造血干细胞是移植到其他器官如骨髓、脾和肺中的粗胎肝细胞制剂中的主要组分，通过Dlk-1选择完全去除，并且Dlk-1纯化的FLSPC仅重新填充肝脏。结论：这是第一个报道从胎儿肝脏中纯化肝干/祖细胞的研究，这些干/祖细胞完全能够重建正常成人肝脏。这代表了开发肝细胞移植治疗临床应用必不可少的方案的重大进展。
Background & Aims: Previously, we showed high-level, long-term liver replacement after transplantation of unfractionated embryonic day (ED) 14 fetal liver stem/progenitor cells (FLSPC). However, for clinical applications, it will be essential to transplant highly enriched cells, while maintaining high repopulation potential. Methods: Dlk-1, a member of the delta-like family of cell surface transmembrane proteins, is highly expressed in human and rodent fetal liver. Dlk-1(+) cells, isolated from ED14 fetal liver using immunomagnetic beads, were examined for their hepatic gene expression profile and characteristic properties in vitro and their proliferative and differentiation potential in vivo after transplantation into normal adult rat liver. Results: Rat ED14 FLSPC were purified to 95% homogeneity and exhibited cell culture and gene expression characteristics expected for hepatic stem/progenitor cells. Rat ED14 FLSPC are alpha-fetoprotein(+)/cytokeratin-19(+) or alpha-fetoprotein(+)/cytokeratin-19(-) and contain all of the normal liver repopulation capacity found in fetal liver. Hematopoietic stem cells, a major component in crude fetal liver cell preparations that engraft in other organs, such as bone marrow, spleen, and lung, are totally removed by Dlk-1 selection, and Dlk-1 purified FLSPC repopulate only the liver. Conclusions: This is the first study reporting purification of hepatic stem/progenitor cells from fetal liver that are fully capable of repopulating the normal adult liver. This represents a major advance toward developing protocols that will be essential for clinical application of liver cell transplantation therapy.