摘要
目的研究人脐带分离的间充质干细胞向神经细胞分化的可能性.方法将剔除动静脉的新鲜人脐带组织切成小块培养,得到贴壁细胞;经传代培养、细胞周期分析、流式细胞检测后,再以不同方案诱导向其神经细胞分化,并以免疫荧光和RT-PCR方法进行鉴定.结果培养5-7d后,有细胞从组织块中游出.细胞传代培养达23代后无明显的形态和增殖能力改变.细胞周期分析表明80%以上的细胞都处于G0～G1期.流式细胞检测表明这些细胞表达CD13、CD29、CD44、CD90、CD105和CD166等MSCs标志物.经神经分化诱导后,部分细胞呈现出与神经元或神经胶质细胞类似的形态;免疫荧光检测表明,第二神经分化诱导方案优于第一方案,其NSE和MBP阳性细胞分别达80.8%±3.9%、4.2%±1.3%,但未能见到GFAP阳性细胞.RT-PCR进一步证实了这些神经标志物的表达.结论人脐带间充质干细胞具有向神经细胞分化的潜能,可作为神经系统疾病细胞移植治疗的备选来源.
Objective To study neural differentiation potential of mesenchymal stem cells (MSCs) from human umbilical cord. Methods After stripping off arteries and veins, the remaining parts of umbilical cord were cut into small sections and plated in 75cm2 flasks. Adherent cells coming out of fragments and were passaged at a dilution of 1:3. Then these cells were proceeded for cell-cycle analysis and fluorescence-activated cell sorting (FACS). Neural induction processes were realized in two different neural induction media, followed by immunocytochemistry analysis and RT-PCR analysis. Results 5-7 days after primary culture, adherent cells came out of fragments and these cells could be maintained for 23 passages without obvious changes in morphology or growth pattern. Doubling time of these cells were estimated about 30 hours and cell cycle analysis revealed that the majority of cells (over 80 percents) stood in G0-G1 phase while a small population of cells was in (S+G2+M) phase. FACS revealed that these cells expressed common markers of MSCs, such as CD13, CD29, CD44, CD90, CD105 and CD166. They also differentiated into NSE positive neural cells with high ratio( 80.8%±3.9%) in the second neural induction medium. Conclusions MSCs from human umbilical cord show an important neural differentiation potential and may represent a novel candidate resource for cell therapy of neurological diseases.
出处
《中华神经外科杂志》
CSCD
北大核心
2005年第7期388-392,共5页
Chinese Journal of Neurosurgery
基金
河北省自然科学基金项目(302508)
