本文關(guān)鍵詞： 脊髓損傷 細(xì)胞培養(yǎng) 移植 慢病毒轉(zhuǎn)染 序貫移植　出處：《安徽醫(yī)科大學(xué)》2017年碩士論文　論文類型：學(xué)位論文

【摘要】：目的:通過(guò)比較由單獨(dú)神經(jīng)干細(xì)胞(NSCs)移植治療大鼠脊髓損傷(SCI)和由骨髓間充質(zhì)干細(xì)胞(BMSCs)與NSCs序貫治療大鼠SCI后大鼠脊髓恢復(fù)情況的不同,研究BMSCs對(duì)大鼠脊髓微環(huán)境的影響以及序貫治療的可行性和優(yōu)勢(shì)。方法:體外分離,培養(yǎng),純化NSCs和BMSCs,慢病毒轉(zhuǎn)染標(biāo)記神經(jīng)干細(xì)胞,改良Allen法制造大鼠脊髓損傷(SCI)模型,將48只大鼠隨機(jī)分為單獨(dú)標(biāo)記神經(jīng)干細(xì)胞移植組(A組,n=24),間充質(zhì)干細(xì)胞+標(biāo)記神經(jīng)干細(xì)胞序貫移植組(B組,n=24)。A組大鼠在T9脊髓損傷后3d經(jīng)尾靜脈注射單細(xì)胞濃度為/101ml6?的POLY-M慢病毒轉(zhuǎn)染后的NSCs條件培養(yǎng)液2ml;B組大鼠在T9脊髓損傷后1d經(jīng)尾靜脈注射單細(xì)胞濃度為/101ml6?的BMSCs條件培養(yǎng)液1ml,脊髓損傷后3d經(jīng)尾靜脈注射單細(xì)胞濃度為/101ml6?的POLY-M慢病毒轉(zhuǎn)染后的NSCs條件培養(yǎng)液2ml。每3只大鼠一組分籠飼養(yǎng),由兩名未參與分組的研究生對(duì)兩組大鼠進(jìn)行BBB評(píng)分,記錄數(shù)據(jù)。分別于移植當(dāng)天,移植后3天、1周、2周,3周、4周、6周、8周各取出A、B組3只大鼠,灌注法取脊髓損傷區(qū)的脊髓組織,福爾馬林固定,制成石蠟切片,行HE染色和MAP-2以及GFAP免疫組化染色,觀察脊髓空洞形成情況,神經(jīng)干細(xì)胞分化生長(zhǎng)情況,以及脊髓膠質(zhì)疤痕大小。結(jié)果:序貫移植組BBB評(píng)分比單獨(dú)神經(jīng)干細(xì)胞移植組高(P0.05);觀察大鼠在治療后1,2,3,4,6,8周時(shí)的神經(jīng)元細(xì)胞數(shù)目,血管生長(zhǎng)情況,膠質(zhì)疤痕以及脊髓空洞大小A,B組之間有顯著性差異(P0.05)。結(jié)論:BMSCs可促進(jìn)了NSCs的有效分化,減少膠質(zhì)化傾向,改善了脊髓的微環(huán)境有利于治療SCI,骨髓間充質(zhì)干細(xì)胞與神經(jīng)干細(xì)胞序貫移植對(duì)大鼠脊髓損傷的療效優(yōu)于單獨(dú)移植神經(jīng)干細(xì)胞。
[Abstract]:Objective: To compare the separate neural stem cells (NSCs) transplantation for the treatment of spinal cord injury (SCI) and bone marrow mesenchymal stem cells (BMSCs) and NSCs sequential treatment of rat SCI rat spinal cord after the recovery of the different effects of BMSCs on rat spinal cord microenvironment and the feasibility of sequential therapy and advantages. Methods: in vitro, separation, purification of NSCs and BMSCs, transfected with lentivirus labeled neural stem cells in rats with spinal cord injury, manufacturing method of modified Allen (SCI) model, 48 rats were randomly divided into separate labeled neural stem cells transplantation group (A group, n= 24), mesenchymal stem cells sequential + labeled neural stem cells transplantation group (group B, n=24).A T9 in rats after spinal cord injury 3D by intravenous injection of single cell concentration is /101ml6? POLY-M lentivirus transfected NSCs conditioned medium 2ml; rats in the B group at T9 after spinal cord injury 1D by caudal vein injection of single cell the concentration of /101ml6? The conditioned medium of BMSCs 1ml, 3D after spinal cord injury by intravenous injection of single cell concentration is /101ml6? POLY-M lentivirus transfected NSCs conditioned medium 2ml. 3 rats each group housed separately by two graduate students did not participate in the group of BBB scores of the two groups of rats were recorded data on the day of transplantation. After transplantation, 3 days, 1 weeks, 2 weeks, 3 weeks, 4 weeks, 6 weeks, 8 weeks each take A, 3 rats in group B, spinal cord, and spinal cord injury zone perfusion method in formalin fixed, paraffin sections were stained with HE and MAP-2 and GFAP immune immunohistochemical staining, syringomyelia formation, differentiation of neural stem cell growth, and spinal cord glial scar size. Results: sequential transplantation group BBB score than single neural stem cell transplantation group (P0.05); the number of cells in the neurons were observed after 1,2,3,4,6,8 weeks of treatment the rats, blood vessel growth, and glial scar The spinal cord cavity size A, there was significant difference between B groups (P0.05). Conclusion: BMSCs can promote the differentiation of NSCs effectively, reduce glial tendency, improve the micro environment of the spinal cord is conducive to the treatment of SCI, bone marrow mesenchymal stem cells and neural stem cells transplantation on the sequential curative effect is better than that of spinal cord injury rats injury transplantation of neural stem cells.

【學(xué)位授予單位】：安徽醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R651.2

【參考文獻(xiàn)】

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本文編號(hào)：1468247