本文關(guān)鍵詞：小鼠肌肉衛(wèi)星細(xì)胞的原代培養(yǎng)和鑒定　出處：《中國(guó)人民解放軍軍醫(yī)進(jìn)修學(xué)院》2007年碩士論文　論文類型：學(xué)位論文

  更多相關(guān)文章： 小鼠 肌肉 衛(wèi)星細(xì)胞 細(xì)胞培養(yǎng)

【摘要】： 目的建立可靠的小鼠肌肉衛(wèi)星細(xì)胞原代培養(yǎng)方法及鑒定技術(shù)。 方法取新生小鼠后肢肌獲得細(xì)胞懸液，經(jīng)Percoll分離，結(jié)合差速貼壁法提純肌肉衛(wèi)星細(xì)胞。倒置顯微鏡觀察，，繪制細(xì)胞生長(zhǎng)曲線，采用結(jié)蛋白、α-肌動(dòng)蛋白免疫細(xì)胞化學(xué)方法鑒定肌肉衛(wèi)星細(xì)胞。 結(jié)果原代骨骼肌衛(wèi)星細(xì)胞在開始分化前呈圓形，通過(guò)細(xì)胞分離、提純后純度為80％以上。在F10培養(yǎng)液內(nèi)肌肉衛(wèi)星細(xì)胞生長(zhǎng)良好，1天后細(xì)胞逐漸貼壁生長(zhǎng)，細(xì)胞較小，分布均勻，多呈圓形；3-6天增長(zhǎng)旺盛；到7-10天可生長(zhǎng)至單層成片約50％～70％；12-14天后肌肉衛(wèi)星細(xì)胞增殖減緩。10-15天的衛(wèi)星細(xì)胞結(jié)蛋白、肌動(dòng)蛋白染色清晰。 結(jié)論使用Percoll分離、差速貼壁法的培養(yǎng)可獲得純度較高的肌肉衛(wèi)星細(xì)胞，而結(jié)蛋白、肌動(dòng)蛋白免疫細(xì)胞化學(xué)染色是鑒定肌衛(wèi)星細(xì)胞的有效方法。
[Abstract]:Objective to establish a reliable primary culture method and identification technique for mouse muscle satellite cells. Methods Cell suspensions were obtained from the hindlimb muscles of newborn mice. The muscle satellite cells were isolated by Percoll and purified by differential adhesion method. The cell growth curve was drawn by inverted microscope and desmin was used. Muscle satellite cells were identified by 偽-actin immunocytochemistry. Results the primary skeletal muscle satellite cells were round before differentiation. The purity of the satellite cells was more than 80% after isolation. The muscle satellite cells grew well in F10 medium. After one day, the cells gradually adhered to the wall, the cells were small, distributed evenly, and most of them were round. The growth was strong in 3-6 days. To 7-10 days can grow to monolayer, about 50%; After 12-14 days, the proliferation of muscle satellite cells slowed. 10-15 days after desmin, actin staining was clear. Conclusion High purity muscle satellite cells can be obtained by Percoll isolation and differential adherence method, and desmin, actin immunocytochemical staining is an effective method for the identification of muscle satellite cells.
【學(xué)位授予單位】：中國(guó)人民解放軍軍醫(yī)進(jìn)修學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2007
【分類號(hào)】：R329.2

【參考文獻(xiàn)】

相關(guān)期刊論文 前7條

1 邵素霞,張雷,趙春芳,鄭力芬,閆蘊(yùn)力,馬洪駿;新生大鼠骨骼肌衛(wèi)星細(xì)胞的原代培養(yǎng)及標(biāo)記方法[J];基礎(chǔ)醫(yī)學(xué)與臨床;2004年01期

2 李志華,宋錦

本文編號(hào)：1360694