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

  更多相關文章： 臍血干細胞 分離 培養(yǎng) 腦出血 移植

【摘要】：研究包括以下兩個部分： 一、臍帶血干細胞的體外分離、培養(yǎng)及鑒定 目的主要采用密度梯度離心法結合直接貼壁法分離人臍帶血中的干細胞，并在體外給予原代及傳代培養(yǎng)，同時觀察細胞形態(tài)，鑒定其增殖能力及是否表達神經干細胞特異性抗原。方法參照文獻[1]的方法，利用干細胞分離液得到臍血干細胞，將其在體外應用DMEM/F12培養(yǎng)基進行原代及傳代培養(yǎng)，采用免疫熒光細胞化學染色方法觀察細胞形態(tài)、鑒定細胞增殖能力及是否表達神經干細胞表面抗原。結果臍血干細胞原代及傳代培養(yǎng)，克隆球中含有表達神經巢蛋白（Nestin）及神經元特異性烯醇化酶（NSE）的陽性細胞，經多次傳代培養(yǎng)后，臍血干細胞可穩(wěn)定增殖，均有一定比例表達Nestin特異性抗原陽性的細胞，且比例無明顯變化。結論實驗證實人臍血干細胞中可分離培養(yǎng)出神經干細胞，反復傳代后部分細胞表達NSE，具有自我更新和增殖的能力，并具有多向分化潛能，從而為臨床神經系統(tǒng)疾病的治療奠定了堅實的基礎。 二、臍血干細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胞再生角度入手，以國內外關于臍帶血干細胞體外誘導、分化成神經干細胞及其在哺乳類動物體內遷移再生等理論研究為基礎，著重研究臍帶血干細胞移植到腦出血后遺癥患者蛛網膜下腔，通過神經功能恢復及腦脊液中細胞生長因子的改變，客觀評價其對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人隨機分為兩組：治療組：將分離出來的臍帶血干細胞經腰椎穿刺部位注入患者蛛網膜下腔，同時給予適當脫水及神經營養(yǎng)藥物等對癥治療。對照組：除了未給予干細胞移植治療外，余治療同治療組。兩組于治療后3、6、9個月神經功能采用斯堪的納維亞卒中量表（Scandinavian Stroke Scale, SSS）進行評分；治療前和治療后1、2、3周分別行腦脊液細胞生長因子檢測。結果1.治療組與對照組在治療前及治療后3、6、9個月采用SSS進行評分，結果顯示治療組明顯高于對照組，P0.05差異有統(tǒng)計學意義。2.臍血干細胞移植組治療后腦脊液細胞生長因子顯著增加，P0.05差異有統(tǒng)計學意義。結論臍血干細胞移植治療腦出血后遺癥取得了一定的療效，為神經系統(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.
【學位授予單位】：內蒙古醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2012
【分類號】：R329

【參考文獻】

相關期刊論文 前10條

1 馮梅;高紅霞;代喜平;盧愛麗;錢彩文;;臍血干細胞移植治療重癥腦癱30例臨床療效觀察[J];中國輸血雜志;2011年07期

2 向靜;王昌銘;王景周;;臍血單個核細胞體外分離和定向分化為神經干細胞的實驗研究[J];重慶醫(yī)學;2007年13期

3 杜冀暉;張鮑虎;李蓉;麥麗文;周蓓;楊佳勇;;人臍血干細胞蛛網膜下腔移植向神經干細胞分化的初步研究[J];國際檢驗醫(yī)學雜志;2008年08期

4 樊建玲;黃河;;骨髓間充質干細胞的免疫學特性及其在造血干細胞移植中的應用[J];國際輸血及血液學雜志;2006年01期

5 許倩,趙連三,王麗春,汪成孝,賀茜,劉麗,吳雄志,于曉輝;人臍帶血間充質干細胞分離培養(yǎng)的技術初探[J];華西醫(yī)學;2005年02期

6 彭紅梅,陳貴安;人類胚胎干細胞體外誘導分化為神經干細胞[J];中國科學C輯:生命科學;2005年03期

7 徐玉;劉靜;王秀麗;王穎;趙平平;郭力;;多發(fā)性硬化治療研究新進展[J];臨床薈萃;2007年15期

8 潘鳳華;李曉波;丁新生;張煒民;張智弘;鄧小萱;姚娟;;人臍血干細胞移植治療大鼠腦缺血的實驗研究[J];臨床神經病學雜志;2006年02期

9 金譽,單根法,鐘z

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