【摘要】：[目的] 驗證大鼠骨髓干細胞(MSCs)體外培養(yǎng)的方法及其生物學特性，研究MSCs的多項分化潛能以及其在體外定向誘導分化為骨髂肌細胞，設(shè)計一組動物模型，分別將誘導后、未誘導的MSCs、陽性對照細胞注射到動物模型體內(nèi)，研究MSCs在體內(nèi)的誘導分化情況及其最終的形態(tài)變化；從而，找到肌肉缺損最佳的修復方法，為臨床治療開拓廣闊的前景。 [方法] 首先培養(yǎng)大鼠骨髓干細胞(MSCs)、大鼠骨骼肌細胞(Mb，，主要用于陽性對照)，將體外培養(yǎng)的MSCs第三代用5—氮雜胞苷誘聯(lián)合MyoD、TGF-β1、IGF-1導分化，用免疫組織化學方法鑒定誘導后的細胞。制做動物模型：取16只10周齡的Wistar大鼠，在其左后小腿脛前肌中部注射無水乙醇0.2ml，使其脛前肌中部變性壞死。右后小腿脛前肌中部注射無菌生理鹽水0.2ml。將16只大鼠模型隨機分成四組，第一組陰性對照，只注射培養(yǎng)基；第二組實驗組，注射MSCs；第三組實驗組，注射誘導后的MSCs；第四組陽性對照組，注射Mb。分別將第三代細胞、誘導后的細胞用BrdU標記，然后收集等待注射。動物模型建立4h后，就可以注射。注射部位雙小腿脛前肌中部。在注射細胞后的第3d、6d、9d和12d分別取材進行形態(tài)觀察和免疫學檢查； [結(jié)果] 大鼠MSCs體外培養(yǎng)生長良好，形態(tài)不規(guī)則，流式細胞儀檢測絕大部分細胞處于G1期(79.4％)，免疫組化檢測CD44陽性，而CD34陰性。大鼠Mb體外培養(yǎng)生長良好，形態(tài)規(guī)則，流式細胞儀檢測絕大部分細胞處于G1期(79.8％)，免疫組化檢測Desmine，Myosin均陽性。經(jīng)誘導后MSCs，形態(tài)多
[Abstract]:[objective] to verify the method and biological characteristics of rat bone marrow stem cell (MSCs) cultured in vitro, to study the differentiation potential of MSCs and its directional differentiation into osteoiliac muscle cells in vitro, and to design a group of animal models. The uninduced MSCs, positive control cells were injected into the animal model to study the differentiation of MSCs in vivo and its final morphological changes, so as to find the best method of repairing muscle defect and open up a broad prospect for clinical treatment. [methods] cultured rat bone marrow stem cell (MSCs),) rat skeletal muscle cells (Mb, was mainly used as positive control). The third generation of MSCs was induced by 5-azacytidine combined with MyoD,TGF- 尾 1IGF-1. The induced cells were identified by immunohistochemical method. Animal model: sixteen 10-week-old Wistar rats were injected with anhydrous ethanol 0.2 ml in the middle of the anterior tibial muscle of the left posterior leg to make the degeneration and necrosis of the middle part of the anterior tibial muscle. The right posterior leg anterior tibial muscle was injected with sterile saline 0.2 ml. Sixteen rats were randomly divided into four groups, the first group was negative control group, the other group was injected with culture medium, the second group was injected with MSCs;, the third group was injected with MSCs;, the fourth group was injected with MSCs; positive control group, and the second group was injected with Mb.. The third generation cells were labeled with BrdU and then collected for injection. The animal model can be injected 4 hours later. The injection site is the middle of the anterior tibial muscle of both legs. Morphological observation and immunological examination were performed on the 3rd day, 6th day and 12th day after injection. [results] Rat MSCs cultured in vitro grew well and had irregular morphology. Flow cytometry showed that most of the cells were in G1 phase (79.4%). Immunohistochemical staining showed that CD44 was positive, but CD34 was negative. Rat Mb grew well in vitro and its morphology was regular. Flow cytometry showed that most of the cells were in G1 phase (79.8%), and Desmine,Myosin was positive by immunohistochemistry. After induction, the morphology of MSCs, was numerous.
【學位授予單位】：山東大學
【學位級別】：碩士
【學位授予年份】：2006
【分類號】：R329.2;R687.2

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