本文關(guān)鍵詞：臍血干細(xì)胞體外分離培養(yǎng)鑒定及用于腦出血后遺癥的療效分析　出處：《內(nèi)蒙古醫(yī)科大學(xué)》2012年碩士論文　論文類型：學(xué)位論文

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【摘要】：研究包括以下兩個(gè)部分： 一、臍帶血干細(xì)胞的體外分離、培養(yǎng)及鑒定 目的主要采用密度梯度離心法結(jié)合直接貼壁法分離人臍帶血中的干細(xì)胞，并在體外給予原代及傳代培養(yǎng)，同時(shí)觀察細(xì)胞形態(tài)，鑒定其增殖能力及是否表達(dá)神經(jīng)干細(xì)胞特異性抗原。方法參照文獻(xiàn)[1]的方法，利用干細(xì)胞分離液得到臍血干細(xì)胞，將其在體外應(yīng)用DMEM/F12培養(yǎng)基進(jìn)行原代及傳代培養(yǎng)，采用免疫熒光細(xì)胞化學(xué)染色方法觀察細(xì)胞形態(tài)、鑒定細(xì)胞增殖能力及是否表達(dá)神經(jīng)干細(xì)胞表面抗原。結(jié)果臍血干細(xì)胞原代及傳代培養(yǎng)，克隆球中含有表達(dá)神經(jīng)巢蛋白（Nestin）及神經(jīng)元特異性烯醇化酶（NSE）的陽性細(xì)胞，經(jīng)多次傳代培養(yǎng)后，臍血干細(xì)胞可穩(wěn)定增殖，均有一定比例表達(dá)Nestin特異性抗原陽性的細(xì)胞，且比例無明顯變化。結(jié)論實(shí)驗(yàn)證實(shí)人臍血干細(xì)胞中可分離培養(yǎng)出神經(jīng)干細(xì)胞，反復(fù)傳代后部分細(xì)胞表達(dá)NSE，具有自我更新和增殖的能力，并具有多向分化潛能，從而為臨床神經(jīng)系統(tǒng)疾病的治療奠定了堅(jiān)實(shí)的基礎(chǔ)。 二、臍血干細(xì)胞移植治療腦出血后遺癥的療效分析 目的本課題主要從腦細(xì)胞再生角度入手，以國內(nèi)外關(guān)于臍帶血干細(xì)胞體外誘導(dǎo)、分化成神經(jīng)干細(xì)胞及其在哺乳類動(dòng)物體內(nèi)遷移再生等理論研究為基礎(chǔ)，著重研究臍帶血干細(xì)胞移植到腦出血后遺癥患者蛛網(wǎng)膜下腔，通過神經(jīng)功能恢復(fù)及腦脊液中細(xì)胞生長因子的改變，客觀評(píng)價(jià)其對(duì)腦出血后遺癥的治療療效，，從而為腦出血的治療探討新的途徑。方法60人隨機(jī)分為兩組：治療組：將分離出來的臍帶血干細(xì)胞經(jīng)腰椎穿刺部位注入患者蛛網(wǎng)膜下腔，同時(shí)給予適當(dāng)脫水及神經(jīng)營養(yǎng)藥物等對(duì)癥治療。對(duì)照組：除了未給予干細(xì)胞移植治療外，余治療同治療組。兩組于治療后3、6、9個(gè)月神經(jīng)功能采用斯堪的納維亞卒中量表（Scandinavian Stroke Scale, SSS）進(jìn)行評(píng)分；治療前和治療后1、2、3周分別行腦脊液細(xì)胞生長因子檢測(cè)。結(jié)果1.治療組與對(duì)照組在治療前及治療后3、6、9個(gè)月采用SSS進(jìn)行評(píng)分，結(jié)果顯示治療組明顯高于對(duì)照組，P0.05差異有統(tǒng)計(jì)學(xué)意義。2.臍血干細(xì)胞移植組治療后腦脊液細(xì)胞生長因子顯著增加，P0.05差異有統(tǒng)計(jì)學(xué)意義。結(jié)論臍血干細(xì)胞移植治療腦出血后遺癥取得了一定的療效，為神經(jīng)系統(tǒng)疾病的治療帶來了新的曙光。
[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.
【學(xué)位授予單位】：內(nèi)蒙古醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2012
【分類號(hào)】：R329

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