細(xì)胞工程論文摘要及總結(jié)-資料下載頁
2025-06-24 00:17本頁面
【正文】 Received 27 March 2006。 accepted 3 July 2006. Available online 18 November 2006. AbstractMouse embryonic stem (ES) cells were cultured on four types of polymer with different surface properties. The polymers were poly(acrylic acid), polyallylamine, gelatin, and poly(2methacryloyloxyethyl phosphorylcholinecomethacrylic acid) (PMAc50), and were coupled with azidophenyl groups and photoimmobilized on conventional polystyrene cellculture dishes. Mouse ES cells were cultured on the immobilized polymer surfaces, and cell morphology, cell growth, staining for alkaline phosphatase, activation of the transcription factor stat3, and expression of the octamerbinding protein 3/4 (Oct3/4) transcription factor and the zinc fingercontaining transcription factor (GATA4) were observed. Morphology and growth rate were significantly affected by the polymer surface properties. The ES cells attached to gelatin or polyallylamine surfaces。 however, colonies formed on the former but not the latter. In addition, significant enhancement of growth was observed on the gelatin surface. In contrast, ES cells aggregated to form an embryoid body on the photoimmobilized poly(acrylic acid) surface and the PMAc50 surface, although cell growth was reduced. Significant enhancement of aggregation of ES cells on the PMAc50 surface was observed in morphology and gene expression analyses.Key words: embryonic stem (ES) cell。 photoimmobilization。 cell culture總結(jié) 胚胎干細(xì)胞（embryonic stem cell, ES）：是胚胎發(fā)育期的胚細(xì)胞，可以人工培養(yǎng)增殖并具有分化成其它類型細(xì)胞的功能。 ES細(xì)胞的形態(tài)學(xué)特征 ES細(xì)胞具有與早期胚胎細(xì)胞相似的形態(tài)結(jié)構(gòu)，細(xì)胞核大，有一個(gè)或幾個(gè)核仁，胞核中多為常染色質(zhì)，胞質(zhì)胞漿少，結(jié)構(gòu)簡單。體外培養(yǎng)時(shí)，細(xì)胞排列緊密，呈集落狀生長。用堿性磷酸酶染色，ES細(xì)胞呈棕紅色，而周圍的成纖維細(xì)胞呈淡黃色。細(xì)胞克隆和周圍存在明顯界限，形成的克隆細(xì)胞彼此界限不清，細(xì)胞表面有折光較強(qiáng)的脂狀小滴。細(xì)胞克隆形態(tài)多樣，多數(shù)呈島狀或巢狀。小鼠ES細(xì)胞的直徑7μm～18μm，豬、牛、羊ES細(xì)胞的顏色較深，直徑12μm～18μm。 從以上論文中可以得知，獲得胚泡方法上的差異不是影響ES細(xì)胞分離克隆的主要影響因素。超排卵的方法可以利用較少的實(shí)驗(yàn)動(dòng)物獲得較多的胚泡,不僅可以節(jié)約實(shí)驗(yàn)成本,而且母鼠受精時(shí)間易于控制,且不影響ES細(xì)胞的克隆率。飼養(yǎng)層的存在有利于胚泡的發(fā)育,胚泡在有飼養(yǎng)層存在時(shí)的48 h貼壁率較無飼養(yǎng)層時(shí)明顯提高,兩者差異有顯著性意義(P005)。在分離培養(yǎng)ES細(xì)胞時(shí)，建議使用新制備的飼養(yǎng)層,以防止ES細(xì)胞在長期體外培養(yǎng)過程中,衰老的飼養(yǎng)層細(xì)胞誘導(dǎo)ES細(xì)胞的體外分化。飼養(yǎng)層細(xì)胞最好是使用3~5代的胎兒成纖維細(xì)胞, 3~5代與原代的胎兒成纖維細(xì)胞,在胚泡的貼壁率、孵化率以及ICM的增殖程度上都無明顯差異。但是原代的胎兒成纖維細(xì)胞的純度低,雜細(xì)胞太多,影響試驗(yàn)的參數(shù)較多,而3~5代的胎兒成纖維細(xì)胞的純度高,可降低試驗(yàn)的影響參數(shù)?？傊?，ES細(xì)胞是一種高度未分化的多潛能干細(xì)胞, 在一定條件下能在體外向各個(gè)胚層分化,目前已廣泛用于體外定向誘導(dǎo)分化的研究。眾多的因素均可以影響ES細(xì)胞的生長,如培養(yǎng)基、血清、各種添加劑、溶液的pH值、溫度、CO2的質(zhì)量、飼養(yǎng)層的種類及狀態(tài)等,任何一方面的不適當(dāng)都可以導(dǎo)致建系的失敗。10 / 10
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