成纤维细胞转录BDNF基因后的条件培养液在体外培养中对神经突起生长的作用  

EFFECTS ON NEURTIE OUTGROWTH IN CONDITIONED MEDIA DERIVERED FROM BDNF GENE-TRANSFERRED FIBROBLAST IN VITRO

作　　者：汪洋[1] 沈馨亚[1] 

机构地区：[1]上海医科大学基础医学院解剖学教研室,上海200032

出　　处：《解剖学杂志》1999年第5期376-380,共5页Chinese Journal of Anatomy

摘　　要：将带有 ＢＤＮＦ基因的成纤维细胞（ＮＩＨ／３Ｔ３）（由上海生化所提供）冻存－７０℃， ６个月后将此工程细胞复苏，放在３７℃、５％ＣＯ_２培养箱中培养扩增，观察形态学并同时收集其培养液，对新生ＳＤ大鼠的脊神经节、脊髓和大脑皮层中的神经元进行条件培养并设对照组。脊神经节植块培养２天后显示ＢＤＮＦ转基因成纤维细胞的条件培养液对促进神经元突起的生长具有良好的活性。对脊髓、大脑皮层中神经元分别进行分散原代培养，１５天后，其突起长度作图像分析（Ｌｅｉｃａ Ｑ５００ ＩＷ）并分别计算出它们的平均长度。脊髓实验组为 １９２． ２０±７５μｍ，对照组为１６０．３７±４８μｍ。大脑皮质实验组为３４７．１４±１０６μｍ，对照组为１２５．６７±４２μｍ。两组的ｔ检验分别是Ｐ＜０．０５。结果显示ＢＤＮＦ转基因成纤维细胞的条件培养液对体外培养神经元突起的生长具有一定的促生长作用。The human fibroblasts were gene-transferred successfully from the BDNF clone by using gene-transferred engineering at Shanghai Research institute of Biochemistry. The BDNF expressing fibroblasts had been stored in -70℃ for six months and then were rewarmed and cultured for recovery and proliferation in 37℃, 5% CO_2 condition. Living observation and morphological recording were also made on the consecutive days. The engineered fibroblasts grew well, and their conditioned media was also collected for the next step. The spinal ganglion, spinal cord and the cerebral cortex of the new-born SD rat were cultured as usual and were divided into two groups: the experimental group with conditioned media mentioned above and the control group. The average length of the neurite outgrowth was analyzed and calculated under Leica Image Analyzer. The results were as follows: The average length of the neurite outgrowth was 192. 20±75 μm in the spinal cord group while 160. 37±48μm in its control group, and 347. 14±106 μm in the cerebral cortex group while 125. 67±42μm in the control group (P<0. 05). We concluded that the conditioned media from BDNF gene-transferred fibroblasts seemed effective in CNS neu- rite outgrowth in vitro, and their morphological features were also well observed in the study.

关 键 词：脑源性 神经生长因子 脊神经节 脊髓 BDNF 

分 类 号：R322.8[医药卫生—人体解剖和组织胚胎学]