CAJ | 학술논문

目的检测体外培养的神经干细胞三种神经营养因子基因的翻译水平和神经生长因子的分泌.方法取孕16 d胚胎SD大鼠皮层细胞,无血清选择性培养液培养神经干细胞,酶联免疫吸附试验(ELISA)法检测第1～7代细胞体外分泌神经生长因子的速度,相对定量实时定量聚合酶链反应(real time-PeR)检测第5～7代细胞脑源性神经营养因子、神经生长因子、神经营养因子-3 mRNA的相对表达量.结果体外培养的神经干细胞在第6代分泌NGF速度最大,(17.80 pg/106细胞·d-1).在第6代BDNF和NT-3基因的相对翻译量最大,在第7代NGF的相对翻译量最大(与对照比值分别为29.446、35.017和16.220).结论神经干细胞在体外翻译、表达并分泌神经营养因子.
목적 검측체외배양적신경간세포삼충신경영양인자기인적번역수평화신경생장인자적분비.방법 취잉16 d배태SD대서피층세포,무혈청선택성배양액배양신경간세포,매련면역흡부시험(ELISA)법검측제1～7대세포체외분비신경생장인자적속도,상대정량실시정량취합매련반응(real time-PeR)검측제5～7대세포뇌원성신경영양인자、신경생장인자、신경영양인자-3 mRNA적상대표체량.결과 체외배양적신경간세포재제6대분비NGF속도최대,(17.80 pg/106세포·d-1).재제6대BDNF화NT-3기인적상대번역량최대,재제7대NGF적상대번역량최대(여대조비치분별위29.446、35.017화16.220).결론 신경간세포재체외번역、표체병분비신경영양인자.
Objective To investigate the relative transcription rate of neurotrophins genes and the expression rate of nerve growth factor (NGF) secreted by neural stem cells. Methods The neural stem cells were harvested from the cortex of E-16 Sprague-Dawley rats,cultured in the neural stem cell-conditioned Dulbecco's Modified Eagle's Medium (DMEM). The secretion rate of NGF in the cells of 1 st -7th passages was determined by ELISA assay, and the relative translation rate of brain-derived neurotrophic factor (BDNF),NGF and neurotrophin-3 (NT-3) in the cells of 5th -7th passages were determined by partial quantitive real-time PCR. Results The peak of NGF secretion rate was at the 6th passage ( 17.80 pg/106 cells·day) in neural stem cells. The peaks of translation rate of BDNF and NT-3 were at the 6th passage (29.446 and 35.107), and of NGF, at the 7th passage ( 16.220). Conclusion Neural stem cells translate,express and secrete neurotrophins in vitro.