自体神经干细胞在兔损伤脊髓内存活并分化的实验效果

Experimental effects of the survival and differentiation of neural stem cells in injured spinal cord of rabbits

目的:将新西兰大白兔骨髓基质细胞体外培养、诱导、分化为神经干细胞,并且移植到兔脊髓横断性损伤模型中,观察移植的细胞是否能够存活及分化。方法:兔骨髓基质细胞取自成年兔的股骨或髂骨,体外培养、诱导、分化为神经干细胞,将兔制成脊髓全横断模型。伤后2周将神经干细胞手术方法直视下移植到受损脊髓缺损处,伤后12周将兔行甲醛灌注留取神经干细胞移植处作为标本,行病理及免疫组化染色。并设立脊髓损伤空白对照组(移植后不行神经干细胞移植)。结果:移植组与对照组相比,移植组移植部位存在大量5-溴-2-脱氧尿苷(bromodeoxyuridine/5-bromo-2-deoxyuridine,BrdU)染色阳性的细胞,表明移植的细胞在体内可以存活;同时,移植组脊髓损伤部位存在神经元特异性烯醇化酶抗体(neuron-specificendase,NSE)、胶质原性纤维酸性蛋白抗体(glialfibrillaryacidicprotein,GFAP)染色阳性的细胞,表明移植的细胞可以分化为具有神经元或胶质细胞特征的细胞。

AIM:To induce and differentiate the bone marrow stromal cells(BMSCs) of New Zealand white rabbits into neural stem cells(NSCs) after culture in vitro,and then transplant the NSCs into the transection models of rabbit spinal cord injury(SCI) so as to observe whether the transplanted cells can survive and differentiate. METHODS:BMSCs derived from femur or ilium of adult rabbits were cultured,and then induced and differentiated into NSCs in vitro,and the complete spinal cord transection models of rabbits were established.Then,at the 2nd week after SCI,the NSCs were transplanted into the defects of injured spinal cord by means of operation under direct vision(transplantation group).The NSCs at the transplantation spot were remained as the specimens after instillation with formaldehyde into the rabbits at week 12 after SCI,and then pathological observation and immunohistochemical staining were performed on the specimens.The rabbits with SCI were conducted as control group(NSCs transplantation was not performed). RESULTS:Compared with the control group,a great number of Bromodeoxy Uridine/5 Bromo 2 Deoxy Uridine(BrdU) positive cells existed at the transplantation spot in the transplantation group,showing that the transplanted cells could survive in vivo.Positive cells of neuron specific endase and glial fibrillary acidic protein also existed at the transplantation spot in the transplantation group,showing that the transplanted cells could be differentiated into neuron like or astrocyte like cells. CONCLUSION:The autologous NSCs in injured spinal cord can survive and be differentiated into neuron like or astrocyte like cells.