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

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。

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

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.