Amit Michal, Winkler Monica E, Menke Sandra, Brüning Eva, Büscher Kristina, Denner Joachim, Haverich Axel, Itskovitz-Eldor Joseph, Martin Ulrich
Department of Obstetrics and Gynecology, Rambam Medical Center, Faculty of Medicine, The Technion, Haifa, Israel.
Stem Cells. 2005 Jun-Jul;23(6):761-71. doi: 10.1634/stemcells.2004-0046.
Until recently, culture and expansion of nondifferentiated human embryonic stem cells (hESCs) depended on coculture with murine embryonic fibroblasts. Because mice are known to harbor a variety of pathogens, such culture conditions implicate the risk of xenozoonoses. Among these pathogens, endogenous retroviruses, including murine leukemia viruses (MuLVs), are of special importance. It is well known that some strains cause pathogenic (e.g., leukemic) effects and that xenotropic, polytropic, and amphotropic MuLVs are able to infect human cells. In view of potential clinical applications of hESC lines, it is therefore imperative to investigate potential infection of hESCs by mouse feeder cell-derived viruses. As a first step towards a comprehensive infection risk assessment, we have analyzed embryonic fibroblasts derived from different mouse strains for expression and release of xenotropic, polytropic, and amphotropic MuLVs. Moreover, several hESC lines have been investigated for expression of specific receptors for xenotropic/polytropic MuLVs, as well as for MuLV infection and expression. Evidence for expression of humantropic MuLVs was found in cultures of mouse embryonic fibroblasts (MEFs). Moreover, expression of specific receptors for xenotropic/ polytropic MuLV on human HEK293 and hESC lines and infection after coculture with an MuLV-producing mink cell line could be demonstrated. In contrast, no evidence of MuLV transmission from MEFs to human HEK293 cells or to the hESC lines I-3, I-6, I-8, and H-9 has been obtained. Our results suggest that recently established hESC lines are free of MuLV infections despite long-term close contact with MEFs.
直到最近，未分化的人类胚胎干细胞（hESCs）的培养和扩增还依赖于与小鼠胚胎成纤维细胞的共培养。由于已知小鼠携带多种病原体，这种培养条件存在异种动物疾病的风险。在这些病原体中，包括鼠白血病病毒（MuLVs）在内的内源性逆转录病毒尤为重要。众所周知，一些毒株会引起致病性（如白血病）效应，而异嗜性、多嗜性和双嗜性MuLVs能够感染人类细胞。鉴于hESC系的潜在临床应用，因此必须研究小鼠饲养层细胞衍生病毒对hESCs的潜在感染。作为全面感染风险评估的第一步，我们分析了来自不同小鼠品系的胚胎成纤维细胞中异嗜性、多嗜性和双嗜性MuLVs的表达和释放情况。此外，还研究了几种hESC系中异嗜性/多嗜性MuLVs特异性受体的表达情况，以及MuLV感染和表达情况。在小鼠胚胎成纤维细胞（MEFs）培养物中发现了嗜人MuLVs表达的证据。此外，还证实了人类HEK293细胞和hESC系中异嗜性/多嗜性MuLV特异性受体的表达，以及与产生MuLV的貂细胞系共培养后的感染情况。相比之下，未获得MuLV从MEFs传播至人类HEK293细胞或hESC系I-3、I-6、I-8和H-9的证据。我们的结果表明，尽管与MEFs长期密切接触，但最近建立的hESC系未感染MuLV。
