大鼠胚胎神经上皮细胞侧脑室移植后存活及分化的实验研究

SURVIVAL AND DIFFERENTIATION OF EMBRYONIC NEUROEPITHELIAL CELLS FOLLOWING TRANSPLANTATION INTO THE LATERAL VENTRICLE OF RATS

目的 观察大鼠胚胎神经上皮细胞同种脑移植后的存活及生长分化状况。 方法 孕 12d大鼠剖腹取胚胎 ,剥离神经管 ,胰酶消化后获取神经管壁上的神经上皮细胞 ,然后植入其父体侧脑室中。分别于移植后 10d、14d、2 1d、2 8d灌注取脑 ,用HE染色及神经元特异烯醇化酶 (NSE)、胶质纤维酸性蛋白 (GFAP)免疫组织化学方法检测胚胎神经上皮细胞移植后的存活及分化状况。 结果 神经上皮细胞侧脑室移植后多贴附于脑室壁形成细胞团块 ,有的随脑脊液循环至第三脑室生长。随时间延长移植物增大。HE染色见脑室内有成团的移植细胞 ;免疫组织化学染色显示移植细胞团内既有NSE 免疫阳性细胞 ,也有GFAP 免疫阳性细胞。移植物周围多为GFAP 免疫阳性细胞。 结论 胚胎神经上皮细胞侧脑室移植后能够贴附脑室壁存活 。

Objective The aim of the study was to observe the survival and differentiation of embryonic neuroepithelial cells in lateral ventricle of rats after transplantation. Methods Embryonic rats(E12d) were obtained from the pregnant Sprague\|Dawley rats;neuroepithelial cells were dissociated from embryonic neural tube and treated with 0\^25% trypsin,then transplanted into the lateral ventricle of father rat.The hosts were anesthetized and transcardially perfused with PBS followed by 4% paraformaldehyde fixative by 10?d,14?d,21?d and 28?d transplantation respectively.Coronal sections of the brain were cut with in cryostat microtome.Using the techniques of histology staining,Nissl staining and immunocytochemistry, survival of neuroepithelial cells transplant and neuro\|specific enolase (NSE) and glial fibrillary acidic protein (GFAP) expression of some stem cells were examined. Results Some grafts were found survival which attached to the ventricle by day 10.Some grafts could migrate into the third ventricle and grew into a mass of cells.Some stem cells were able to differentiate into pyramidal cells.By immunocytochemistry,a mass of NSE\|positive cells and GFAP\|positive cells could be detected in the core of graft region.Around intracerebral grafts,some astrocytes showed immunoreactive for glial fibrillary acidic protein.Conclusion\ These findings demonstrate that embryonic stem cells dissociated from neural tube can survive and differentiate into neurons and astrocytes.