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Establishment of an immortal cynomolgus macaque fibroblast cell line for propagation of cynomolgus macaque cytomegalovirus (CyCMV)

机译:建立永生食蟹猕猴成纤维细胞系以繁殖食蟹猕猴巨细胞病毒(CyCMV)

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摘要

Cynomolgus macaques are widely used as an animal model in biomedical research. We have established an immortalized cynomolgus macaque fibroblast cell line (MSF-T) by transducing primary dermal fibroblasts isolated from a 13-year-old male cynomolgus macaque with a retrovirus vector expressing human telomerase reverse transcriptase (hTERT). The MSF-T cells showed increased telomerase enzyme activity and reached over 200 in vitro passages compared to the non-transduced dermal fibroblasts, which reached senescence after 43 passages. The MSF-T cell line is free of mycoplasma contamination and is permissive to the newly identified cynomolgus macaque cytomegalovirus (CyCMV). CyCMV productively infects MSF-T cells and induces down-regulation of MHC class I expression. The MSF-T cell line will be extremely useful for the propagation of CyCMV and other cynomolgus herspesviruses in host-derived fibroblast cells, allowing for the retention of host-specific viral genes. Moreover, this cell line will be beneficial for many in vitro experiments related to this animal model.
机译:食蟹猕猴被广泛用作生物医学研究中的动物模型。我们通过用表达人端粒酶逆转录酶(hTERT)的逆转录病毒载体转导从13岁的雄性食蟹猕猴分离的初级真皮成纤维细胞,建立了永生的食蟹猕猴成纤维细胞系(MSF-T)。与未转导的真皮成纤维细胞相比,MSF-T细胞显示出增加的端粒酶活性,并在体外传代了200多次,后者在43传代后达到了衰老。 MSF-T细胞系不含支原体污染,并且允许新近识别的食蟹猕猴巨细胞病毒(CyCMV)。 CyCMV有效感染MSF-T细胞并诱导MHC I类表达下调。 MSF-T细胞系对于CyCMV和其他食蟹猕猴疱疹病毒在宿主来源的成纤维细胞中的繁殖非常有用,可保留宿主特异性病毒基因。而且,该细胞系对于与该动物模型有关的许多体外实验将是有益的。

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