c—kit在肠神经节细胞发育过程中的作用  

The role of c-kit on the development of enteric neurons

作　　者：吴晓娟[1] 冯杰雄[1] 魏明发[1] 柴成伟[1] 郑帅玉[1] 高贺云[1] 王果[1] 

机构地区：[1]武汉华中科技大学同济医学院附属同济医院小儿外科,430030

出　　处：《中华小儿外科杂志》2009年第11期785-788,共4页Chinese Journal of Pediatric Surgery

基　　金：湖北省自然科学基金资助项目（编号：2007ABA092）

摘　　要：目的初步探讨c-kit在肠神经节细胞发育中的作用。方法体外培养大鼠肠神经节细胞及骨髓间充质干细胞（BMSCs），并在GDNF与胎肠培养基（FGCM）培养的条件下诱导BMSCs分化为肠神经节细胞。免疫组织化学方法鉴定原代培养的肠神经节细胞神经丝蛋白（neurofilament，NF）的表达，用血管活性肽（vasoactive intestinal peptide，VIP）及NF鉴定BMSCs诱导分化的肠神经节细胞。RT-PCR分别检测原代肠神经节细胞、BMSCs以及诱导之肠神经节细胞的c—kit的表达。结果免疫组化结果显示纯化后的肠神经节细胞表达NF，流式细胞学检测第四代后的BMSCs高表达CD90，而不表达CD45，诱导后的肠神经节细胞可表达VIP及NF，而其余两组则不表达。RT-PCR结果示原代肠神经节细胞及BMSCs不表达c-kit，诱导后的肠神经节细胞可表达c-kit。结论在体外能成功培养大鼠肠神经节细胞及BMSCs并纯化，BMSCs可被诱导为肠神经节细胞并表达肠神经递质。在肠神经系统的发育过程中，c-kit可能在某一阶段起到一定作用，在肠神经细胞逐渐发育成熟时接受到某些信号而关闭。Objective To investigate the possible role of c-kit on the development of enteric neurons. Methods Enteric neurons and bone marrow stromal cells （BMSCs） were harvested and cultured from rats. At passage 4,BMSCs were induced into enteric neurons by GDNF of 10ng/ml in fetal gut condition medium （FGCM） for 7 days. Primary enteric neurons were identified by neurofilament （NF） and induced enteric neurons were identified by vasoactive intestinal peptide （VIP） and NF with immunohistochemical assay. The expression of c-kit was detected by RT-PCR. Results After purification, primary enteric neurons showed expression of NF. BMSCs at passage 4 were CD90 positive and CD45 negative detected by flow cytometry. After 7 days of induction, induced enteric neurons expressed VIP,while BMSCs were VIP negative in the control group. Primary enteric neurons and BMSCs didn＇t express c-kit mRNA. The expression of ckit mRNA of induced enteric neurons increased highly after induction. Conclusions Enteric neurons and BMSCs can be harvested and cultured in vitro,and BMSCs can be induced to differentiate into enteric neurons. C-kit may play a role at a certain stage of development of enteric nervous system and be blocked after receiving a signal of mature enteric neurons.

关 键 词：肠神经细胞 细胞分化 C-KIT 

分 类 号：R686[医药卫生—骨科学]