Fetal Liver Stem/Progenitor Cell Transplantation: A Model to Study Tissue Mass Replacement and Cell-Based Therapies.
Fetal Liver Stem/Progenitor Cell Transplantation: A Model to Study Tissue Mass Replacement and Cell-Based Therapies.
复制标题
胎儿肝干/祖细胞移植：研究组织块替代和细胞疗法的模型。
DOI：
10.1007/978-1-4939-6506-9_7
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发表时间：
2017
期刊：
影响因子：
--
通讯作者：
Oertel,Michael
中科院分区：
文献类型：
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作者：
Yovchev,MladenI;Oertel,Michael
肝移植是终末期肝病患者的唯一治疗方法。然而，供体器官稀缺是主要限制，因此，迫切需要替代策略。成功的基于细胞的治疗的最终目标是移植细胞能够有效地植入和重建受损的肝脏肿块。为了评估移植细胞的再繁殖能力，有必要确定它们的特性，并利用成熟的细胞移植模型研究移植供体细胞在肝脏微环境中替代组织块的机制(S)。到目前为止，大鼠胎肝干/祖细胞是重建接近正常的肝脏和晚期纤维化/肝硬化肝脏微环境的最有效的细胞群，因此，可以用来开发未来工程潜在供体细胞的策略，这些策略将有助于在肝细胞治疗中的临床应用。本方案描述了从DPPIV+F344或F344-TG(EGFP)F455/Rrrc大鼠的ED14/15胎肝分离的上皮干/祖细胞的分离，免疫组织化学染色方法检测E-钙粘素阳性上皮细胞的分离，并将其移植到不同的DPPIV-肝微环境中(接近正常，逆行处理，治疗，和TAA诱导的肝纤维化/肝硬变)，以及跟踪移植细胞在受体肝脏中命运的检测方法(参见图。1)。
Liver transplantation is the only therapeutic treatment for patients with end-stage liver diseases. However, donor organ scarcity is the major limitation, and therefore, alternative strategies are urgently needed. The ultimate goal for successful cell-based therapies is the ability of transplanted cells to efficiently engraft and reconstitute injured liver mass. To evaluate the repopulation capacity of transplanted cells, it is essential to identify their specific characteristics, as well as to study the mechanism(s) Through which transplanted donor cells replace tissue mass in hepatic microenvironments, using well-established cell transplantation models. To date, rat fetal liver stem/progenitor cells represent the most efficient cell population to reconstitute the near-normal liver and the liver microenvironment with advanced fibrosis/cirrhosis, and therefore, can be used for developing strategies in engineering potential donor cells in the future that will be useful for clinical application in hepatic cell therapy.The present protocol describes the isolation of epithelial stem/progenitor cells derived from ED14/15 fetal livers of DPPIV+F344 or F344-Tg(EGFP) F455/Rrrc rats, the immunohistochemical staining method to detect E-cadherin-positive epithelial cells within unfractionated cell isolates, their transplantation into different DPPIV–liver microenvironments (near-normal, retrorsine-treated, and TAA-induced fibrotic/cirrhotic liver), as well as detection methods to follow the fate of transplanted cells in the recipient liver (seeFig. 1).