转染绿色荧光蛋白的神经干细胞移植于半横断脊髓损伤处的体内外分化研究

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.

Differentiate of neural stem cell labelled by green fluorescent protein in rat semi - transected spinal cord in vitro and in vivo

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.