摘要
目的:比较白细胞介素1α与胶质细胞源神经营养因子不同组合条件下骨髓基质细胞分化情况,探讨骨髓基质细胞诱导分化为神经干细胞及多巴胺能神经元的分化条件。方法:2005-03/09在解放军第一军医大学珠江医院全军神经医学研究所进行。①实验分组:实验分为空白对照组;胶质细胞源神经营养因子+白细胞介素1α组,后者又分为10μg/L+100ng/L,10μg/L+200ng/L,20μg/L+100ng/L,20μg/L+200ng/L,1000μg/L+100ng/L组。②细胞模型制作:用全骨髓法培养骨髓基质细胞。③骨髓基质细胞以5×108L-1接种,悬浮于本实验室配制的神经干细胞培养基中。空白对照组:血清终浓度为体积分数为0.1的胎牛血清。细胞接种48h后,胶质细胞源神经营养因子+白细胞介素1α各组分别加入相应质量浓度的胶质细胞源神经营养因子+白细胞介素1α。④应用Olympus光学显微镜观察细胞抗-神经巢蛋白和D2受体阳性染色阳性细胞数目计数。结果:①加入胶质细胞源神经营养因子、白细胞介素1α增殖培养48h可见细胞分裂相。②诱导6d,部分细胞有神经巢蛋白成分表达。③3周测出DA受体D2检测阳性,胶质细胞源神经营养因子+白细胞介素1α10μg/L+100ng/L组细胞诱导分化率显著高于10μg/L+200ng/L,20μg/L+100ng/L,20μg/L+200ng/L,1000μg/L+100ng/L组及空白对照组犤依次为:(11.70±0.55)%,(3.62±0.78)%,(4.14±0.41)%,(3.3±0.63)%,(4.92±0.34)%,(1.61±0.68)%,P<0.05或0.01犦。结论:骨髓基质细胞在体外培养条件下,经过胶质细胞源神经营养因子、白细胞介素1α诱导分化作用,配合使用高浓度(体积分数为0.1)血清可获得巢蛋白阳性的神经前体细胞及表达D2受体阳性的多巴胺能神经元;10μg/L+100ng/L为最佳诱导分化质量浓度。
AIM： To compare the differentiation of bone marrow stromal cells（BMSCs） under the condition of different combinations of interleukin-let and gliacyte-derived neurotrophic factor so as to investigate the condition of BMSCs inducing and differentiating into neural stem cells and dopaminergic neuron. METHODS： This experiment was conducted at the Institute of Neuromedicine, Zhujiang Hospital, First Military Medical University of Chinese PLA from March to September 2005. ① Experiment grouping： The experiment was divided into blank control group; gliacyte-derived neurotrophic factor＋interleukin let group, and the latter was subdivided into 10μg/L＋100ng/L, 10μg/L＋200 ng/L,20 μg/L＋100 ng/L,20 μg/L＋ 200 ng/L and 1 000 μg/L＋100 ng/L groups. ②Preparation of cell model： BMSCs were cultured with general bone marrow methods.③ BMSCs were inoculated with the density of 5×10^8L^-1 and suspended in the culture medium of neural stem cells made in our laboratory . Blank control group： the final concentration of serum was 0.1 fetal bovine serum. After the cells were inoculated for 48 hours, gliacyte-derived neurotrophic factor＋interleukin let was added into the corresponding quality concentration groups. ④Tbe number of positive cells of cell anti- nerve nidogen and D2 receptor positive staining with Olympus optical microscope. RESULTS： ①Cell division appeared when gliacyte-derived neurotrophic factor＋interleukin let were added for proliferative culture for 48 hours . ②Some cells had the expression of the component of nerve nidogen after induction for 6 days. ③ At week 3, DA receptor D2 was positive . The induction and differentiation rate of gliacyte-derived neurotrophic factor＋ interleukin 1α 10μg/L＋100 ng/L group was significantly higher than that in the 10μg/L＋200 ng/L,20μg/L＋100 ng/L,20μg/L＋200 ng/L, 1 000 μg/L＋100 ng/L group as well as blank control group lit was （11.70±0.55）%, （ 3.62±0.78 ）%, （4.14±0.41 ）%, （ 3.3±0.63 ）%, （4.92±0.34）%
出处
《中国临床康复》
CSCD
北大核心
2006年第9期31-33,i0002,共4页
Chinese Journal of Clinical Rehabilitation
