人Muse细胞体外诱导为神经前体细胞的研究

Study on Induction of Human Muse Cells into Neural Precursor Cells in vitro

探讨成人骨髓来源的Muse细胞体外诱导为神经前体细胞的技术方法。从健康成人骨髓中分离出骨髓基质细胞(h BMSCs),利用流式细胞分选技术从中筛选出Muse细胞,采取免疫荧光细胞化学染色和q PCR技术鉴定Muse细胞的多能干细胞特性;通过神经诱导培养基体外诱导Muse细胞分化为神经前体细胞(Muse-NPCs),采用免疫荧光细胞化学染色和q PCR技术鉴定其神经干细胞标志物表达情况。从h BMSCs中分选出约0.58%的Muse细胞,Muse细胞表达多能干细胞标志物SSEA-3、Nanog和Oct4,其mRNA表达水平均高于h BMSCs(P<0.01);诱导后的Muse-NPCs表达神经干细胞标志物Nestin、βIII-tubulin,其mRNA表达水平均高于h BMSCs及Muse细胞(P<0.01)。从成人骨髓中成功分离出具有多能干细胞特性的Muse细胞,用体外诱导形成Muse-NPCs,可为今后细胞治疗修复神经损伤提供新的种子细胞。

The work explored approaches of inducing Muse cells derived from the adult bone marrow into neural precursor cells in vitro. Human bone marrow stromal cells（ h BMSCs） were separated and cultured from the healthy adult bone marrow. Then Muse cells were collected using the flow cytometry sorting. In order to observe the characteristics of the pluripotent stem cells,Muse cells were analyzed using immunocytochemical staining and q PCR assay. Afterward,neural inducing culture media was used to induce Muse cells into neural precursor cells（ Muse-NPCs）. The immunocytochemical staining and q PCR assay were used to observe the expression of the markers specific for the neural stem cell in Muse cells. Muse cells accounting for about 0. 58% of h BMSCs were obtained that expressing the markers of the pluripotent stem cells： SSEA- 3,Nanog and Oct4. Muse cells also showed significantly higher levels of mRNA of these markers comparing to h BMSCs（ P 〈 0. 01）. The induced Muse-NPCs expressed the markers of neural stem cells including Nestin and βIII-tubulin. Additionally,the levels of mRNA of these markers in Muse-NPCs was significantly higher than those in h BMSCs and Muse cells（ P 〈 0. 01）. In conclusion,Muse cells were successfully separated from adult bone marrow and then induced into Muse-NPCs in vitro,providing a new type of seed cells for the cell therapies in the nervous system.