CAJ | 학술논문

目的观察转染绿色荧光蛋白(GFP)的大鼠脊髓神经干细胞移植于半横断脊髓损伤处的体内外分化情况.方法将表达GFP的慢病毒载体转染胎鼠脊髓神经干细胞,体外用10%胎牛血清诱导分化.转染后的神经干细胞与PLGA支架移植于大鼠半横断脊髓损伤处,术后1个月和3个月取材,行GFAP、NF和CNP免疫荧光染色.结果转染GFP的神经干细胞球表达强烈的绿色荧光,体外分化可见GFAP/GFP、NF/GFP和CNP/GFP双阳性细胞,GFAP/GFP双阳性细胞明显多于其他两种.移植后3个月,GFP阳性细胞在脊髓内明显减少,可见少数GFAP/GFP和CNP/GFP舣阳性细胞,未见NF/GFP双阳性细胞.结论转染GFP的神经干细胞可在体外增殖和分化,但大部分分化成胶质细胞.移植于急性期脊髓损伤处的神经干细胞不被诱导分化成神经元样细胞,可被诱导分化成神经胶质细胞.
목적 관찰전염록색형광단백(GFP)적대서척수신경간세포이식우반횡단척수손상처적체내외분화정황.방법 장표체GFP적만병독재체전염태서척수신경간세포,체외용10%태우혈청유도분화.전염후적신경간세포여PLGA지가이식우대서반횡단척수손상처,술후1개월화3개월취재,행GFAP、NF화CNP면역형광염색.결과 전염GFP적신경간세포구표체강렬적록색형광,체외분화가견GFAP/GFP、NF/GFP화CNP/GFP쌍양성세포,GFAP/GFP쌍양성세포명현다우기타량충.이식후3개월,GFP양성세포재척수내명현감소,가견소수GFAP/GFP화CNP/GFP의양성세포,미견NF/GFP쌍양성세포.결론 전염GFP적신경간세포가재체외증식화분화,단대부분분화성효질세포.이식우급성기척수손상처적신경간세포불피유도분화성신경원양세포,가피유도분화성신경효질세포.
Objective To investigate the differentiation of rat spinal cord neural stem cells(NSCs) labelled by green fluorescent protein in vitro and in rat semi - transected spinal cord injury. Method NSCs from the spinal cord of embryonic rat were cultured in vitro, and transfected by lentiviral vectors expressing green fluorescent protein ( GFP) . The transfected NSCs supported by poly lactide -co - glycolide acid (PLGA) were implanted into rat semi - transected spinal cord. 1 and 3 months later, the biological characteristics and differentiated activity were identified by immunofluorescence and observed under fluoroscope. Results GFAP/GFP?NF/GFP and CNP/GFP double - positive cells could be seen in vitro. In 3 months after transplantation, GFP positive cells could be found in spinal cord, but the number was markedly less than in 1 month. GFAP/GFP and CNP/GFP double - positive cells could be seen, and no GFAP/GFP cells under fluorescence microscope. Conclusions NSCs labelled by GFP have the capability of self - renewing and multipotential differentiation in vitro, they are not induced to differentiate into like -neuron cell in acute spinal cord injury,but glial cells.