体外诱导的皮肤源性神经干细胞在大鼠受损伤脊髓内的存活、分化和迁移
首发时间:2006-01-11
摘要:目的:探讨经体外诱导的皮肤源性神经干细胞能否在大鼠脊髓半横断损伤处存活、迁移和分化。 方法:应用转染绿色荧光蛋白基因新生大鼠的皮肤在体外经分离细胞、培养、诱导增殖以及用免疫细胞化学染色鉴定神经干细胞等过程, 然后将诱导产生的皮肤源性神经干细胞移植入大鼠脊髓半横断损伤处,30d和60d后用免疫细胞化学染色法观察移植细胞存活、迁移和分化。 结果:皮肤分离细胞在培养10d时,呈悬浮生长的细胞已增殖成许多的细胞球, 并呈nestin免疫细胞化学阳性染色, 表明是神经干细胞球。在体内可观察到脊髓损伤处有许多带有绿色荧光的移植皮肤源性神经干细胞,有些还迁移到较远的宿主脊髓组织内。存活的移植细胞有些呈现nestin阳性、MAP2阳性及GFAP阳性。 结论:经体外诱导的皮肤源性神经干细胞能够在受损伤的大鼠脊髓内存活、迁移并分化为神经元样细胞和星形胶质样细胞。
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The survival, differentiation and migration of skin-derived neural stem cells in rat injured spinal cord
Abstract:Objective To investigate whether the skin-derived neural stem cells inducted by the culture could survive, differentiate and migrate in the lesioned site of rat spinal cord. Methods The skin of new born rat trans-gene of green fluorescence protein (GFP) was applied to be dissociated into cells, cultured and inducted to proliferate in vitro. Skin-derived neural stem cells were identified by immunocytochemistry staining. Then, skin-derived neural stem cells inducted by the culture were transplanted into the lesioned site of rat spinal cord hemisection. 30 days and 60 days after the operation, the survival, migration and differentiation of transplanted cells were observed by immunocytochemistry staining. Results 10 days at cultured skin-dissociated cells, many cell-spheres had been formed by the proliferation of suspensive growing cells. These cell-spheres showed nestin positive staining of immunocytochemistry. It suggestes that the cell-spheres are neurospheres. At vivo, many transplanted skin-derived neural stem cells with GFP were observed in the lesioned area of spinal cord. Some transplanted cells migrated into host spinal cord tissue far away from the lesioned area. Some transplanted surviving cells showed nestin, MAP2 and GFAP positive staining of immunocytochemistry separately. Conclusion Skin-derived neural stem cells inducted by the culture may survive, migrate and differentiate into neuron-like cells and astrocyte-like cells in injured spinal cord of rat.
Keywords: Skin stem cell Neural stem cell Spinal cord injury Survival Differentiation
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