摘要
目的 观察神经营养因子-3（NT-3）在体内外能否诱导脊髓源性神经干细胞分化为胆碱能神经元。方法 从孕龄15d的胚胎sD大鼠脊髓组织中分离获得脊髓源性神经干细胞。体外诱导分化研究：实验组在培养脊髓源性神经干细胞时，去除表皮生长因子和碱性成纤维细胞生长因子加入NT-3进行诱导；对照组则单纯培养脊髓源性神经干细胞。体内诱导分化研究：实验组将去除表皮生长因子和碱性成纤维细胞生长因子加入NT-3的脊髓源性神经干细胞混合悬液移植于成年SD大鼠切断胫神经远端，对照组则移植未加NT-3脊髓源性神经干细胞悬液。细胞免疫荧光化学法鉴定分化结果。结果 经NT-3诱导的脊髓源性神经干细胞用细胞免疫荧光化学法鉴定，体内外培养均可检测到胆碱乙酰基转移酶阳性细胞，而对照组则未检测到胆碱乙酰基转移酶阳性细胞。结论 NT-3在体内外均可诱导脊髓源性神经干细胞分化为胆碱能神经元。
Objective To investigate whether neurotrophin-3 （NT-3） can induce in vivo and in vitro the spinal cord-derived neural stem cells to differentiate into cholinergic neurons. Methods The spinal cord-derived neural stem cells of 15-day SD fetal rats were harvested and cultivated in serum-free media. For in vitro investigation, the spinal cord-derived neural stem cells were cultured after NT-3 was added and EGF and bFGF were removed in the experimental group, while the spinal cord-derived neural stem cells were simply cultured without addition of NT-3 to the culture media in the control group. For in vivo investigation, the suspension of the neural stem cells with NT-3 and without EGF or bFGF was injected into the distal stump of transected tibial nerve in adult SD rats in the experimental group, while only the suspension of neural stem cells was injected into the distal stump of transected tibial nerve in the control group. The results of differentiation were identified by cellular immunohistochemistry. Results The spinal cord-derived neural stem cells differentiated into choline acetyltransferase-positive cells when induced by NT-3 in vivo and in vitro. But in the control groups, no choline acetyltransferase-positive cells were detected. Conclusion NT-3 can induce the spinal cord-derived neural stem cells to differentiate into cholinergic neurons in vivo and vitro.
出处
《中华创伤骨科杂志》
CAS
CSCD
2007年第6期546-549,共4页
Chinese Journal of Orthopaedic Trauma
基金
国家自然科学基金（30271308）
上海市科委资助项目（04JC14024）
