目的对成人骨髓源性神经干细胞(Md-NSCs)中重要抑癌基因p53、Rb1及p16进行突变检测,旨在从抑癌基因方面研究Md-NSCs的致瘤性问题。方法从正常成人志愿者获取成人骨髓基质细胞(hMSCs),于体外诱导培养获得Md-NSCs,应用PCR-DNA测序技术,PCR扩增Md-NSCs中p53基因第5-9外显子、Rb1基因第19-21外显子以及p16基因第1-2外显子,并对扩增片断进行DNA测序。结果Md-NSCs可由hMSCs于体外诱导培养简捷获取,Md-NSCs中p53基因第5-9外显子、Rb1基因第19-21外显子以及p16基因第1-2外显子的序列均与野生型一致,无发现突变现象。结论hMSCs体外诱导培养分化形成的Md-NSCs中未发现重要抑癌基因的常见突变现象,保持了遗传学的稳定性,为其体内移植的潜在致瘤性评价提供了参考依据。
Objective To investigate the tumorigenicity of human adult bone marrow-derived neural stem cells (Md-NSCs) by mutation detection of the three important tumor suppressor genes p53,Rbl and p16 in these cells. Methods Human adult marrow stromal cells (bMSCs) were obtained from adult normal volunteers and induction culture into Md-NSCs in vitro,the mutation detection of the exons 5 -9 of p53 gene,the exons 19- 21 of Rbl gene and the exons 1 -2 of p16 gene in Md-NSCs were carried out by polymerase chain reaction and DNA sequencing subsequently. Results Md-NSCs could be efficiently generated from hMSCs by induction culture in vitro,the sequences of the exons 5 -8 of p53 gene,exon 19-21 of Rbl gene and exons 1 -2 of p16 gene in Md-NSCs were all consistent with the wide types,without any mutation found. Conclusion No common mutations of important tumor suppressor genes could be found in Md-NSCs,which provided useful information for assessing the long-term tumorigenicity of these cells upon autologons transplantation.