Abstract

Classical Swine Fever (CSF) is an extremely infectious and deadly disease of pigs and wild boars caused by the CSF virus (CSFV) which is a member of the Pestivirus genus and the family Flaviviridae. This study was designed to detect the permissibility and replication of CSFV in mesenchymal stem cells (MSCs) monolayer derived from Porcine Wharton’s jelly. Porcine Wharton’s jelly MSCs (pWJ-MSCs) were ex vivo expanded and propagated for more than 81 generations and third passage pWJ-MSCs were characterized as per standard criteria i.e., growth characteristics, trilineage differentiation potential and molecular characterization for pluripotency and stem cell surface markers. Porcine WJ tissue samples found negative for CSFV by RT-PCR test were processed further for the isolation of pWJ-MSCs and CSFV was propagated over the characterized pWJ-MSCs monolayer. No cytopathic effect was observed, which was consistent with non-cytopathic nature of CSFV. The replication of CSFV in pWJ-MSCs was affirmed by RT-PCR and demonstration of viral antigen in the cytoplasm of virus infected cells by immuno-staining technique. In total, three different CSFV isolates were propagated in pWJ-MSCs. Primary pWJ-MSCs permitted CSFV replication to good titer. To the best of our information, this is the first ever report of isolation of CSFV in pWJ-MSCs.

摘要

经典猪瘟 (CSF) 是猪和野猪的一种极具传染性和致命的疾病，由 CSF 病毒 (CSFV) 引起，CSFV 是瘟病毒属和黄病毒科的成员。本研究旨在检测源自猪沃顿氏胶的间充质干细胞 (MSCs) 单层中 CSFV 的允许性和复制。Porcine Wharton's 果冻 MSCs (pWJ-MSCs) 是离体的扩增和繁殖超过81代和第三代pWJ-MSC根据标准标准进行表征，即生长特征、三系分化潜能和多能性和干细胞表面标志物的分子表征。通过 RT-PCR 测试发现猪 WJ 组织样本对 CSFV 呈阴性，进一步处理以分离 pWJ-MSCs，并且 CSFV 在表征的 pWJ-MSCs 单层上繁殖。未观察到细胞病变效应，这与 CSFV 的非细胞病变性质一致。RT-PCR证实了CSFV在pWJ-MSCs中的复制，并通过免疫染色技术在病毒感染细胞的细胞质中证实了病毒抗原。总共有三种不同的 CSFV 分离株在 pWJ-MSCs 中繁殖。初级 pWJ-MSCs 允许 CSFV 复制到良好的滴度。