人神经干细胞的分离、克隆和动物脑内移植及转基因表达

目的分离和克隆人神经干细胞,并在体外和体内分析其生物学特征.方法我们联合采用四步法从人胚胎前脑分离制备多潜能神经干细胞,并使用重组腺病毒相关病毒载体(rAAV)将LacZ基因和胶质细胞起源的神经营养因子(GDNF)基因转移到神经干细胞.结果二株人神经干细胞被成功建立.这些克隆化后的神经干细胞在细胞培养中和移植到新生小鼠脑内后能发育分化成神经元、少枝胶质细胞和星形胶质细胞.在rAAV转导基因后,神经干细胞可在体外和体内表达转基因产物.结论这种具有转基因表达能力的神经干细胞为神经系统疾病的进一步治疗研究提供了有潜在价值的细胞资源.

Isolation and cloning of human neural stem cells, intracerebral grafting and transgene expression in mice.

Objective To isolate, clone and characterize human neural stem cells in vitro and in vivo. Methods We combined four procedures for isolation and perpetuating multipotent neural stem cells from human embryonic forebrain. Recomminant adeno-associated viruses(rAAV) were used to mediate LacZ and GDNF gene transduction into neural stem cells. Results Two human neural stem cell line were established. These neural stem cell clones gave rise to neurons, astrocytes, and oligodendrocytes in culture and in the newborn mouse brain following transplantation. After gene transduction by rAAV, the neural stem cells demonstrated the capability of expressing foreign transgenes in vitro and in vivo. Conclusions The human neural stem cells with transgene expression ability provides a potential cell source for therapy and research of neural diaeases.