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  本文關(guān)鍵詞：大鼠肌肉衛(wèi)星細(xì)胞的分離培養(yǎng)鑒定與誘導(dǎo)分化　出處：《內(nèi)蒙古大學(xué)》2010年碩士論文　論文類型：學(xué)位論文

  更多相關(guān)文章： 大鼠 肌肉衛(wèi)星細(xì)胞 成肌誘導(dǎo) 成脂誘導(dǎo) fat-1

【摘要】： 骨骼肌肌肉控制著脊椎動物的精確的運(yùn)動。成體骨骼肌的組成單元是肌纖維。肌纖維是由一群處于有絲分裂間期的成肌細(xì)胞組成的合胞體。而未分化的肌肉衛(wèi)星細(xì)胞參與了合胞體的修復(fù)和維持。 肌肉衛(wèi)星細(xì)胞是未分化的單核肌源性細(xì)胞群落,在哺乳動物、爬行動物、鳥類、兩棲動物的骨骼肌內(nèi)都能發(fā)現(xiàn)。1961年在蛙的肌肉中首先發(fā)現(xiàn)了衛(wèi)星細(xì)胞的存在。電鏡觀察提示衛(wèi)星細(xì)胞位于骨骼肌纖維的肌細(xì)胞膜與基底膜之間,其核較大、胞漿少,細(xì)胞器數(shù)量少。這些特征說明衛(wèi)星細(xì)胞處于有絲分裂靜止期,其轉(zhuǎn)錄活性低于肌纖維。 由于能夠從病人骨骼肌中獲得并在體外大量擴(kuò)增,肌肉衛(wèi)星細(xì)胞可以作為細(xì)胞移植的資源,并且已經(jīng)應(yīng)用于骨骼肌和心肌的重建。盡管發(fā)現(xiàn)衛(wèi)星細(xì)胞的存在已經(jīng)有50多年的歷史了,但分離鑒定肌肉衛(wèi)星細(xì)胞的方法仍然具有不穩(wěn)定性。 本研究使用兩步酶消化和差速貼壁相結(jié)合的方法分離大鼠肌肉衛(wèi)星細(xì)胞。Desmin免疫細(xì)胞化學(xué)染色、Myf5及Myodl的RT-PCR法證明了所分離的細(xì)胞是肌肉衛(wèi)星細(xì)胞。成肌和成脂誘導(dǎo)試驗證明了所分離的肌肉衛(wèi)星細(xì)胞具有多分化潛能。 主要研究結(jié)果如下： 1.通過兩步酶消化和差速貼壁相結(jié)合的方法分離得到了肌肉細(xì)胞系。Desmin免疫細(xì)胞化學(xué)染色、Myf5及Myodl的RT-PCR法證明了所分離的細(xì)胞是肌肉衛(wèi)星細(xì)胞。 2.分離得到的肌肉衛(wèi)星細(xì)胞能夠向成肌細(xì)胞和脂肪細(xì)胞分化的潛能,具有一定的多能性。 3. DCA處理促進(jìn)了肌肉衛(wèi)星細(xì)胞的增殖并激活了WNT/beta-Catenin信號通路。
[Abstract]:Skeletal muscle muscles control the precise movement of vertebrates. Adult skeletal muscles are composed of muscle fibers. Muscle fibers are syncytial cells consisting of a group of myoblasts in the mitotic phase. Undifferentiated muscle satellites. Cells are involved in the repair and maintenance of syncytial bodies. Muscle satellite cells are undifferentiated mononuclear myogenic cell communities in mammals, reptiles, and birds. In 1961, satellite cells were first found in frog muscle. Electron microscopic observation indicated that satellite cells were located between muscle cell membrane and basement membrane of skeletal muscle fiber. These characteristics indicate that the satellite cells are in mitotic quiescent phase and their transcriptional activity is lower than that of muscle fibers. Muscle satellite cells can be used as a resource for cell transplantation because they can be obtained from patients' skeletal muscles and expanded in vitro. Although the existence of satellite cells has been discovered for more than 50 years, the method of isolation and identification of muscle satellite cells is still unstable. In this study, two steps of enzyme digestion and differential adhesion were used to isolate rat muscle satellite cells. Desmin immunocytochemical staining was used. The RT-PCR method of Myf5 and Myodl proved that the isolated cells were muscle satellite cells, and the myoblast and adipogenic induction tests showed that the isolated muscle satellite cells had multi-differentiation potential. The main findings are as follows: 1. The muscle cell line. Desmin immunocytochemical staining was obtained by two-step enzyme digestion and differential adhesion. The RT-PCR method of Myf5 and Myodl proved that the isolated cells were muscle satellite cells. 2. The isolated muscle satellite cells have the potential to differentiate into myoblast and adipocytes, and have certain pluripotency. 3. DCA treatment promoted the proliferation of muscle satellite cells and activated the WNT/beta-Catenin signaling pathway.
【學(xué)位授予單位】：內(nèi)蒙古大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2010
【分類號】：R329

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本文編號：1356374