摘要
目的 利用银杏内酯B（GB）诱导大鼠骨髓间充质干细胞（BMSC）分化成熟,研究其分化的神经样细胞的电生理特征及免疫学特征.为临床治疗神经系统免疫性疾病提供最合适种子细胞.方法 应用倒置相差显微镜对GB诱导的细胞进行多个时间点观察其细胞形态学改变,并对诱导后细胞进行免疫荧光染色;应用膜片钳技术,采取全细胞记录方式,对由GB诱导的大鼠BMSC进行诱导前及诱导后多个时间点的电生理测定.结果 免疫荧光染色阳性细胞：CHAT：（34.8％,n=12）;GFAP：（31.6％,n=12）;NSE：（21.3％,n=12）;膜电位随着时间的变化：未分化细胞的峰值电流（1.45 ±0.15PA/PF,n=10）,诱导后的神经元样细胞（2.18 ±0.14 PA/PF,n=12）变化显著（P＜0.05）.结论 大鼠BMSC经过GB诱导后免疫特性及膜电位均发生改变,显示诱导后细胞有神经干细胞的免疫学特点,可以作为一种调节神经免疫力及修复力的种子细胞.
Objective After using Ginkgolide B （GB） to induce rat bone marrow-derived mesenchy- mal stem cell（BMSC） differentiation and maturation, the electrophysiological and immunological properties of neuron-like cells were explored. These cells may provide the seed ceils for the clinical treatment of autoimmune disease of nervous system, such as Guillain-Barre syndrome. Method The cells induced by GB at different times were observed with a inverted phase contrast microscope and subjected to immunofluorescence staining. Patch clamp recording technique using a whole-cell recording mode was adopted to record the electrophysiologi- cal properties of the BMSC before and after the induction by GB. Results After GB induction, there was 34. 8 % （ n = 12 ） of choline-acetyltransferase-positive （CHAT） cells, 31.6% （ n = 12 ） of glial-fibrillary-acidie- protein（GFAP） cells, and 21. 3% （ n = 12） of neuron-specific-enolase （NSE） cells. Membrane potential changed over time; The peak current of undifferentiated cells was 1.45 ±0.15 PA/PF（ n = 10）. After the in- duction by GB, the peak current of neuron ceils was 2.18 ±0.14 PA/PF （ n = 12） , which was significantly different compared to that of undifferentiated cells （ P 〈 0.05 ）. Conclusion Both of the immunological features and membrane potential of BMSCs in rats induced by GB have changed, showing that the induced cells contain immunological characteristics of neural stem cells. Therefore, these cells may have the potential to be the seed cells for adjusting the nerve immunity and repairment capability.
出处
《国际免疫学杂志》
CAS
2014年第5期444-448,共5页
International Journal of Immunology
基金
济宁市科技课题支助（lc2012020）
