Department of Gastroenterology, Xinhua Hospital, College of Medicine, Shanghai Jiao Tong University, Shanghai, China.
J Gene Med. 2010 Mar;12(3):244-54. doi: 10.1002/jgm.1439.
beta(2)m(-)/Thy1(+) bone marrow-derived hepatocyte stem cells (BDHSCs) isolated from the bone marrow of cholestatic rats by magnetic bead cell sorting consistently express characteristics of both stem and liver cells. These stem cells may be good vehicles for gene transfer. Administration of exogenous hepatocyte growth factor (HGF) may be potentially useful for the treatment of liver fibrosis. Because lentiviral vectors integrate stably into the host-cell genome of nondividing and dividing cells, it may efficiently transfect beta(2)m(-)/Thy1(+) BDHSCs in vitro and secrete high-level HGF consistently. Transplantation of beta(2)m(-)/Thy1(+) BDHSCs transduced with lentiviral vectors containing the HGF gene may reduce liver fibrosis in rats.
Lentiviral vectors expressing HGF were constructed and used to transduce beta(2)m(-)/Thy1(+) BDHSCs sorted from cholestatic rats in vitro. Transduction efficiency was evaluated and then these cells were transplanted into rats through the portal vein. Liver function as well as histological and immunohistochemical examinations were carried out to assess the therapeutic efficacy on liver fibrosis.
We demonstrated that high-level exogenous HGF was detected in supernatants after beta(2)m(-)/Thy1(+) BDHSCs were transfected with lentiviral vectors expressing HGF. Transplantation of transduced beta(2)m(-)/Thy1(+) BDHSCs significantly enhanced liver function and attenuated liver fibrosis in vivo.
The present study indicates that transplantation of beta(2)m(-)/Thy1(+) BDHSCs overexpressing the HGF gene may offer a novel approach for promoting liver function and reverse liver fibrosis.
通过磁珠细胞分选从胆汁淤积大鼠骨髓中分离的β(2)m(-)/Thy1(+)骨髓源性肝干细胞（BDHSCs）持续表达干细胞和肝细胞的特征。这些干细胞可能是基因转移的良好载体。外源性肝细胞生长因子（HGF）的给药可能对肝纤维化的治疗有用。由于慢病毒载体稳定地整合到非分裂和分裂细胞的宿主细胞基因组中，因此它可以有效地在体外转染β(2)m(-)/Thy1(+)BDHSCs，并持续分泌高水平的 HGF。转导 HGF 基因的慢病毒载体的β(2)m(-)/Thy1(+)BDHSCs 移植可能会减轻大鼠的肝纤维化。
构建表达 HGF 的慢病毒载体并用于体外转染从胆汁淤积大鼠中分选的β(2)m(-)/Thy1(+)BDHSCs。评估转导效率，然后通过门静脉将这些细胞移植到大鼠体内。进行肝功能以及组织学和免疫组织化学检查，以评估对肝纤维化的治疗效果。
我们证明了β(2)m(-)/Thy1(+)BDHSCs 转染表达 HGF 的慢病毒载体后，上清液中检测到高水平的外源性 HGF。转导的β(2)m(-)/Thy1(+)BDHSCs 移植显著增强了体内肝功能并减轻了肝纤维化。
本研究表明，过表达 HGF 基因的β(2)m(-)/Thy1(+)BDHSCs 移植可能为促进肝功能和逆转肝纤维化提供一种新方法。
