CAJ | 학술논문

背景:干细胞移植是神经损伤修复领域的研究热点,目前多数研究集中于神经干细胞和骨髓干细胞移植研究,而脐带间充质干细胞相关研究目前国内刚刚起步,后者较前者具有来源广泛,伦理限制少等优势. 目的:构建脑源性神经营养因子真核表达载体,并将其转染至人脐带间充质干细胞中,建立稳定表达脑源性神经营养因子的细胞模型. 方法:应用反转录 PCR 获得脑源性神经营养因子的基因序列,然后将脑源性神经营养因子的基因序列插入到 pcDNAⅢ载体的多克隆位点,构建脑源性神经营养因子蛋白表达载体,并转染脐带间充质干细胞;Western Blot 体外检测携带脑源性神经营养因子基因的脐带间充质干细胞的脑源性神经营养因子蛋白表达水平;取培养好的 PC12细胞和胎鼠皮质神经元加入含脑源性神经营养因子的脐带间充质干细胞培养基,检测脑源性神经营养因子蛋白的生物活性.结果与结论:成功构建脑源性神经营养因子真核表达载体,该载体能在脐带间充质干细胞中高水平表达脑源性神经营养因子蛋白,表达的脑源性神经营养因子蛋白体外检测具有生物活性.可见脐带间充质干细胞有可能作为脑源性神经营养因子真核表达载体用于神经损伤修复研究.
배경:간세포이식시신경손상수복영역적연구열점,목전다수연구집중우신경간세포화골수간세포이식연구,이제대간충질간세포상관연구목전국내강강기보,후자교전자구유래원엄범,윤리한제소등우세. 목적:구건뇌원성신경영양인자진핵표체재체,병장기전염지인제대간충질간세포중,건립은정표체뇌원성신경영양인자적세포모형. 방법:응용반전록 PCR 획득뇌원성신경영양인자적기인서렬,연후장뇌원성신경영양인자적기인서렬삽입도 pcDNAⅢ재체적다극륭위점,구건뇌원성신경영양인자단백표체재체,병전염제대간충질간세포;Western Blot 체외검측휴대뇌원성신경영양인자기인적제대간충질간세포적뇌원성신경영양인자단백표체수평;취배양호적 PC12세포화태서피질신경원가입함뇌원성신경영양인자적제대간충질간세포배양기,검측뇌원성신경영양인자단백적생물활성.결과여결론:성공구건뇌원성신경영양인자진핵표체재체,해재체능재제대간충질간세포중고수평표체뇌원성신경영양인자단백,표체적뇌원성신경영양인자단백체외검측구유생물활성.가견제대간충질간세포유가능작위뇌원성신경영양인자진핵표체재체용우신경손상수복연구.
BACKGROUND: Stem cel s transplantation is the research hot spot in the area of nerve injury repair. At present, most researches have focused on neural stem cel s and bone marrow stem cel s transplantation, and there are rare reports on the research of umbilical cord mesenchymal stem cel s, but the umbilical cord mesenchymal stem cel s have the advantages of rich source and little ethical restrictions. @@@@OBJECTIVE: To construct an eukaryotic expression vector expressing brain derived neurotrophic factor, and to transfect it into the umbilical cord mesenchymal stem cel s and establish a cel model of brain derived neurotrophic factor. @@@@METHODS: Firstly, the c-DNA of brain derived neurotrophic factor was gained by reverse transcription-PCR and then inserted into multi-clone site of pcDNAⅢ vector to construct the expression vector of brain derived neurotrophic factor pcDNAⅢ. Then the vector was used to transtect umbilical cord mesenchymal stem cel s. At last, the expression level of brain derived neurotrophic factor in the umbilical cord mesenchymal stem cel s was tested by Western blot; the cultured PC12 cel s and the embryo rat pal ium neurons were added into the umbilical cord mesenchymal stem cel s containing brain derived neurotrophic factor to test the biological activity of brain derived neurotrophic factor. @@@@RESULTS AND CONCLUSION: The eukaryotic expression vector expressing brain derived neurotrophic factor was successful y constructed, and this vector could highly express the brain derived neurotrophic factor protein in umbilical cord mesenchymal stem cel s. In vitro testing showed the brain derived neurotrophic factor protein had the biological activity. It indicates that the umbilical cord mesenchymal stem cel s can be used as the eukaryotic expression vector of brain derived neurotrophic factor for research on the nerve injury repair.