摘要
目的 :体外诱导大鼠骨髓间质干细胞 (MSC)分化为神经细胞。方法 :SD大鼠股骨骨髓细胞体外扩增。用肾上腺素、去甲肾上腺素和异丙肾上腺素注射液分别加入无血清L -DMEM诱导MSC分化为神经细胞。免疫细胞化学鉴定有神经元烯醇化酶 (NSE)、神经干细胞标志物巢蛋白 (nestin)和胶质纤维酸性蛋白 (GFAP)表达。结果 :大鼠骨髓间质干细胞在体外扩增 5 - 2 2代 ,对照组不加任何诱导剂 ,有 5 3%的神经样细胞。加入肾上腺素、去甲肾上腺素和异丙肾上腺素诱导 1- 5h ,约 70 %MSC形态转变为典型的神经样细胞。免疫细胞化学显示诱导出的神经样细胞NSE、nestin、GFAP表达阳性。继续培养 5d后对照组神经样细胞逐渐凋亡。肾上腺素类各组虽存活 6d ,但细胞正常形态改变 ,部分细胞死亡漂浮。一瓶MSC在正常培养至第 7代时 ,自发出现约 5 0 %的神经细胞 ,传至第 13代 ,有约 6 0 %的神经细胞 (6 6 .5 %± 6 .4 % )。结论 :骨髓本身可能存在着神经干细胞。大鼠骨髓间质干细胞用肾上腺素类诱导可分化为多种形态的神经细胞。
AIM:To investigate the differentiation from rat mesenchymal stem cells (rMSC) into neuron-like cells. METHODS:rMSC were separated from femur marrow and expanded in L-DMEM culture medium supplemented with 10% FSC. rMSC were induced to differentiate into neurons with L-DMEM/adrenaline,L-DMEM/noradrenaline and L-DMEM/isoprenaline, respectively. Meanwhile, rMSC were cultured in L-DMEM in control group. Nestin, neuron-specific enclose (NSE), glial fibrillary acidic protein (GFAP) were detected by immunocytochemistry. RESULTS: rMSC were expanded as undifferentiated cells in culture from 5 to 22 passages, indicating their differentiated capacity. Simple method induced rMSC to exhibit a neuronal phenotype, expressing positive NSE,nestin, and GFAP, at 5 hours in all group. The undifferentiating cells (control group 53.1%±4.3%), and differentiating cells (treated group: adrenaline 74.7%±2.6%; noradrenaline 75.9%±2.4%; isoprenaline 72.1%±4.4%), expressed characteristics of various neuronal cells, from 5 hours to 6 days. There were neuron-like cells in rMSC cultured in L-DMEM/10%FBS from 7 to 13 passage(66.5%±6.4%). CONCLUSION: It suggests that rat neural stem cells (rNSC) exist in bone marrow, rMSC can be differentiated into various neural cells with adrenaline hormones in vitro.
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2004年第5期733-736,共4页
Chinese Journal of Pathophysiology
基金
国家科技部 86 3计划资助项目(2 0 0 2AA2 0 4 0 11)
