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目的探索骨髓间充质干细胞(BMSCs)源性神经细胞对C6胶质瘤的趋向性。方法阿尔法最低必需培养基(α-MEM)、神经细胞生长添加剂B27、碱性成纤维细胞生长因子(bFGF)、表皮细胞生长因子(EGF)和全反式维甲酸(ATRA)诱导BMSCs向神经细胞分化。体外通过Transwell系统共培养BMSCs源性神经细胞与C6胶质瘤细胞培养基,HE染色分析穿过聚醋酸纸膜微孔的细胞数量;体内在C6胶质瘤模型中移植5-溴脱氧尿嘧啶核苷(BrdU)标记的BMSCs源性神经细胞,免疫组化分析移植细胞的迁移能力。结果条件培养基诱导BMSCs分化的细胞表达神经元核抗原(NeuN)为47%±0.08%,高分子量神经丝蛋白(NF-H)为45%±0.07%及部分巢蛋白(Nestin)为10%±0.04%,体外Transwell结果显示BMSCs源性神经细胞透过聚碳酸酯膜微孔的数量明显高于对照组(P<0.05),体内移植结果显示BMSCs源性神经细胞与BMSCs对胶质瘤的迁移无明显差异(P>0.05)并呈现时间特异性(P<0.01)。结论 BMSCs源性神经细胞对C6胶质瘤有明显的趋向性。
Objective To explore the tendency of bone marrow mesenchymal stem cells (BMSCs) derived neurons to C6 glioma. Methods Alpha-MEM, B27, bFGF, AFP and all-trans retinoic acid (ATRA) were used to induce neural stem cells Differentiation. BMSCs-derived neurons and C6 glioma cell culture medium were co-cultured with Transwell system in vitro. HE staining was used to analyze the number of cells passing through the microporous cellulose acetate membrane. In vivo, C6 glioma model was transplanted with 5-bromodeoxyuridine (BrdU) labeled BMSCs-derived neurons, and the migration ability of the transplanted cells was analyzed by immunohistochemistry. Results The conditioned medium induced BMSCs expressed NeuN of 47% ± 0.08%, high-molecular-weight neurofilament protein (NF-H) of 45% ± 0.07% and partial nestin of 10% ± 0.04%. Transwell results showed that the number of BMSCs-derived neurons passing through the polycarbonate membrane was significantly higher than that of the control group (P <0.05). Transplantation results showed that BMSCs-derived neurons and BMSCs There was no significant difference in migration (P> 0.05) and showed time specificity (P <0.01). Conclusion BMSCs-derived neurons have obvious tendency toward C6 glioma.