HSV-1感染大鼠骨髓间充质干细胞及形成潜伏感染的初步研究

在体外培养大鼠骨髓间充质干细胞(BMSCs),观察单纯疱疹病毒1型感染骨髓间充质干细胞情况.分离并鉴定BMSCs;HSV-1感染BMSCs,观察细胞病变(CPE);建立BMSCs的HSV-1潜伏感染模型.提取总DNA,PCR法扩增BMSCs内的HSV-1特异性片段,检测HSV-1感染BMSCs及潜伏感染.结果显示骨髓间充质干细胞经14d诱导后,碱性磷酸酶含量增高、形成钙结节,表现出成骨细胞特性.HSV-1感染BMSCs,出现典型的CPE,PCR法证实BMSCs内存在HSV-1的特异性片段.HSV-1潜伏感染的BMSCs,未出现明显的CPE,细胞传至7代,仍可测到HSV-1的基因片段,表明BMSCs有可能形成HSV-1的潜伏感染.大鼠骨髓间充质干细胞在体外可以向成骨细胞方向分化,可作为组织工程学的种子细胞.HSV-1可以在体外感染骨髓间充质干细胞并有形成潜伏感染的趋势.

Infection and Latency of HSV-1 on Rat Bone Marrow Mesenchymal Stem Cells in Vitro

Abstract: To culture bone marrow mesenchymal stem cells(BMSCs) of rat in vitro and observe HSV-1 infection on BMSCs,BMSCs were separated from the bone marrow and identified by alizarin red staining and detection of ALP.The morphology of HSV-1 infected BMSCs and the CPE were observed.The total DNA was extracted from HSV-1 infected BMSCs and the desired specific gene fragment of 477bp of HSV-1 was amplified by PCR.Results showed that after BMSCs were induced by mineral-fluid for 14 days,the ALP level was increased and the nodule calcification was formed.The induced BMSCs were manifested to have the characteristics of osteoblasts.CPE couldn't be found in HSV-1 latently infected BMSCs but the 477bp gene fragment was still detectable.HSV-1 could establish latent infection in BMSCs after 7 passages.This study indicated that rat BMSCs could be induced to differentiate into osteoblasts in vitro,therefore they can be used as the seed cells for the tissue engineering.HSV-1 can infect rat BMSCs and develop the latent infection in vitro.