脐血干细胞体外分离培养鉴定及用于脑出血后遗症的疗效分析

发布时间：2018-01-16 15:06

本文关键词：脐血干细胞体外分离培养鉴定及用于脑出血后遗症的疗效分析　出处：《内蒙古医科大学》2012年硕士论文　论文类型：学位论文

【摘要】：研究包括以下两个部分：一、脐带血干细胞的体外分离、培养及鉴定目的主要采用密度梯度离心法结合直接贴壁法分离人脐带血中的干细胞，并在体外给予原代及传代培养，同时观察细胞形态，鉴定其增殖能力及是否表达神经干细胞特异性抗原。方法参照文献[1]的方法，利用干细胞分离液得到脐血干细胞，将其在体外应用DMEM/F12培养基进行原代及传代培养，采用免疫荧光细胞化学染色方法观察细胞形态、鉴定细胞增殖能力及是否表达神经干细胞表面抗原。结果脐血干细胞原代及传代培养，克隆球中含有表达神经巢蛋白（Nestin）及神经元特异性烯醇化酶（NSE）的阳性细胞，经多次传代培养后，脐血干细胞可稳定增殖，均有一定比例表达Nestin特异性抗原阳性的细胞，且比例无明显变化。结论实验证实人脐血干细胞中可分离培养出神经干细胞，反复传代后部分细胞表达NSE，具有自我更新和增殖的能力，并具有多向分化潜能，从而为临床神经系统疾病的治疗奠定了坚实的基础。二、脐血干细胞移植治疗脑出血后遗症的疗效分析目的本课题主要从脑细胞再生角度入手，以国内外关于脐带血干细胞体外诱导、分化成神经干细胞及其在哺乳类动物体内迁移再生等理论研究为基础，着重研究脐带血干细胞移植到脑出血后遗症患者蛛网膜下腔，通过神经功能恢复及脑脊液中细胞生长因子的改变，客观评价其对脑出血后遗症的治疗疗效，，从而为脑出血的治疗探讨新的途径。方法60人随机分为两组：治疗组：将分离出来的脐带血干细胞经腰椎穿刺部位注入患者蛛网膜下腔，同时给予适当脱水及神经营养药物等对症治疗。对照组：除了未给予干细胞移植治疗外，余治疗同治疗组。两组于治疗后3、6、9个月神经功能采用斯堪的纳维亚卒中量表（Scandinavian Stroke Scale, SSS）进行评分；治疗前和治疗后1、2、3周分别行脑脊液细胞生长因子检测。结果1.治疗组与对照组在治疗前及治疗后3、6、9个月采用SSS进行评分，结果显示治疗组明显高于对照组，P0.05差异有统计学意义。2.脐血干细胞移植组治疗后脑脊液细胞生长因子显著增加，P0.05差异有统计学意义。结论脐血干细胞移植治疗脑出血后遗症取得了一定的疗效，为神经系统疾病的治疗带来了新的曙光。
[Abstract]:The study consists of the following two parts: 1. Isolation, culture and identification of umbilical cord blood stem cells in vitro Objective to isolate stem cells from human umbilical cord blood by density gradient centrifugation combined with direct adherent method. To identify its proliferative ability and whether it expresses neural stem cell specific antigens. [1. The umbilical cord blood stem cells were obtained from stem cell isolate and cultured in DMEM/F12 medium in vitro. The cell morphology was observed by immunofluorescence cytochemical staining, and the proliferation and expression of neural stem cell surface antigen were identified. Results the primary culture and culture of umbilical cord blood stem cells were carried out. Positive cells expressing nestin (Nestin) and neuron-specific enolase (NSE) were found in the cloned spheres. After several passages, cord blood stem cells could proliferate stably. There was no significant change in the proportion of Nestin specific antigen positive cells. Conclusion the neural stem cells can be isolated and cultured from human umbilical cord blood stem cells. After repeated passage, some cells express NSEs, which have the ability of self-renewal and proliferation, and have the potential of multi-differentiation, thus laying a solid foundation for the treatment of clinical nervous system diseases. Second, the therapeutic effect of umbilical cord blood stem cell transplantation on the sequelae of intracerebral hemorrhage Objective to study the neural stem cells (NSCs) induced by umbilical cord blood stem cells in vitro and their migration and regeneration in mammalian animals from the point of view of brain cell regeneration. To study the transplantation of umbilical cord blood stem cells to the subarachnoid space of patients with cerebral hemorrhage sequelae, and to evaluate objectively the therapeutic effect of cord blood stem cells transplantation on the sequelae of cerebral hemorrhage through the recovery of nerve function and the change of cell growth factor in cerebrospinal fluid. Methods 60 patients were randomly divided into two groups: the treatment group: the isolated umbilical cord blood stem cells were injected into the subarachnoid space of the patients by lumbar puncture. At the same time, appropriate dehydration and neurotrophic drugs were given to the symptomatic treatment. The control group: in addition to the stem cell transplantation treatment, the remaining treatment group. The neurological function was evaluated by Scandinavian Stroke scale (SSS) for 9 months. Cerebrospinal fluid growth factor (CSF) was measured before treatment and 3 weeks after treatment. Results 1.The SSS score was used in the treatment group and the control group before and after 3 months and 9 months after treatment. 2. The results showed that the treatment group was significantly higher than the control group P0.05 the difference was statistically significant. 2. Cord blood stem cell transplantation group after treatment of cerebrospinal fluid cell growth factor significantly increased. Conclusion umbilical cord blood stem cell transplantation in the treatment of intracerebral hemorrhage sequelae has a certain effect, which brings a new dawn for the treatment of nervous system diseases.
【学位授予单位】：内蒙古医科大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R329

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