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目的观察骨髓胚胎样干细胞体内再生dystrophin的可行性。方法采用SSP-PCR方法鉴定mdx小鼠的基因型。分离扩增人骨髓胚胎样干细胞,采用DIR标记后,注射细胞到mdx小鼠腹股沟三角皮下,采用活体成像法观察植入细胞的存活情况,采用免疫荧光染色法检测dystrophin的表达。结果杂合子鼠交配可以产生3个基因型的子代鼠,采用SSP-PCR可以鉴定出mdx小鼠基因型;采用无血清培养基可从骨髓中分离到表达SSEA-4和FZD-9的胚胎样干细胞,可在注射细胞的mdx小鼠离体后肢肌肉检测到明显的荧光信号,并在肌肉组织中检测到人dystrophin的表达。结论人骨髓胚胎样干细胞可在mdx小鼠体内存活并表达dystrophin。
Objective To observe the feasibility of regenerating dystrophin in bone marrow-derived embryonic stem cells. Methods The genotypes of mdx mice were identified by SSP-PCR. The human bone marrow-derived embryonic stem cells were isolated and expanded. After labeling with DIR, the cells were injected into the groin triangle of mdx mice. The survival of the implanted cells was observed by live imaging method. The expression of dystrophin was detected by immunofluorescence staining. Results The heterozygous offspring mice could produce three genotypes of offspring. The genotypes of mdx mice were identified by SSP-PCR. The embryos expressing SSEA-4 and FZD-9 were isolated from bone marrow using serum-free medium Like cells, a clear fluorescence signal was detected in isolated hindlimb muscle of mdx mice injected with cells, and the expression of human dystrophin was detected in muscle tissue. Conclusion Human bone marrow-derived embryonic stem cells can survive and express dystrophin in mdx mice.