免疫磁珠法分离纯化胚胎大鼠脊髓源运动神经元的方法探讨  

EXPERIMENTAL RESEARCH OF SEPARATION AND PURIFICATION OF EMBRYONIC RAT SPINAL MOTONEURONS BY MAGNETIC ACTIVATED CELL SORTING

作　　者：王菲[1] 王春芳[1] 李鹏飞[1] 蔚洪恩[1] 韩树峰[2] 李承罡[3] 

机构地区：[1]山西医科大学医学实验中心,太原030001 [2]山西医科大学第一医院骨科,太原030001 [3]山西医科大学第二医院骨科,太原030001

出　　处：《中国组织化学与细胞化学杂志》2009年第4期422-425,共4页Chinese Journal of Histochemistry and Cytochemistry

基　　金：国家自然科学基金资助(30672114);山西省自然科学基金(2009011057-1)

摘　　要：目的探讨应用免疫磁珠法分选脊髓源性运动神经元的实验条件,为研究运动神经元的特性,培养和移植创造有利条件。方法取孕16天胚胎大鼠的脊髓腹侧组织,制备成单细胞悬液,应用免疫磁珠分选系统,在无血清限定性培养基条件下获得相对纯化的运动神经元。应用运动神经元特异的胆碱乙酰转移酶(ChAT)抗体对培养细胞进行免疫细胞化学鉴定。结果分离纯化的细胞在体外可以存活,经免疫荧光检测,ChAT阳性率可达95%。结论本实验提供了一种有效的纯化脊髓运动神经元的方法,纯化所得的运动神经元可用于进一步的运动神经元的后续实验研究。Objective To explore the experimental conditions of sorting spinal motoneurons by mag netic activated cell sorting （MACS）, and to create a favorable condition for studying the characteristics, culture and transplantation of motoneurons. Methods The ventral spinal cord cell suspensions from 16-day rat fetus were isolated, and the motoneurons were purified by MACS and the culture system of a serumfree medium. Spinal motoneurons were identified by immunostaining with special polyclonal antibody to choline acetyltransferase （CHAT） . Results The present experiment revealed that the isolated and puried cells might live in vitro. After the culture of about one week, more than 95% cells were ChAT-positive by immunofluorescent staining. Conclusion This study suggests a purification method of spinal motoneurons, which ensures the cells to be satisfactorily used in further study of motoneurons.

关 键 词：脊髓 运动神经元 免疫磁珠分选 纯化 大鼠 

分 类 号：R322.812[医药卫生—人体解剖和组织胚胎学]