麻省理工懷特黑德研究所等處的研究人員發(fā)現(xiàn)了一個能提高人類胚胎干細(xì)胞,,以及誘導(dǎo)多能干細(xì)胞iPS三倍數(shù)量的新方法,,而且這種方法還無需一般培養(yǎng)方法中都需要的小鼠飼養(yǎng)細(xì)胞,。這一研究成果公布在美國《國家科學(xué)院院刊》(PNAS)雜志上,。
胚胎干細(xì)胞和iPS細(xì)胞由于其杰出的再生能力,,吸引了眾多科學(xué)家,,這些多能干細(xì)胞是許多疾病的新希望,,比如帕金森癥,,阿茲海默癥等,也是篩選信號的新方向,,同時這些細(xì)胞還能為糖尿病,,神經(jīng)或其它組織損傷提供移植治療細(xì)胞。但是這些技術(shù)應(yīng)用需要百萬,,甚至千萬的細(xì)胞,,目前的培養(yǎng)方法無法達(dá)到這一需求。
在這篇文章中,,研究人員僅僅通過紫外線照射了常用的聚苯乙烯實驗平板,,就提高了人類胚胎干細(xì)胞,和iPS細(xì)胞生長速度,,細(xì)胞數(shù)目增長了三倍,,而且還不需要小鼠飼養(yǎng)細(xì)胞。文章的作者,,哈佛-麻省理工學(xué)院衛(wèi)生科學(xué)與技術(shù)部副教授Daniel Anderson表示,,"聚苯乙烯是實驗室最常見的培養(yǎng)表面,這一小小的改動就能替代小鼠飼養(yǎng)層,,相信這將會應(yīng)用到許多方面,。"
利用小鼠成纖維細(xì)胞作飼養(yǎng)層是胚胎干細(xì)胞分離培養(yǎng)和傳代的關(guān)鍵技術(shù)之一,而這種方法會傳播病毒,,或者小鼠大分子,,污染人類干細(xì)胞。為了提高干細(xì)胞培養(yǎng)速度,,以及避免污染物風(fēng)險,這一研究組的成果努力發(fā)掘新方法,,他們在去年的一篇Nature Materials文章中,,報道了利用人工高分子基質(zhì)(含有大量碳?xì)浠衔?,以及酯基)大幅度提高了?xì)胞的生長速度,雖然這一方法很奏效,,但是由于這些聚合物很難合成不同的細(xì)胞培養(yǎng)容器,,因此無法應(yīng)用到大規(guī)模生產(chǎn)中。
而最新的這篇文章在之前研究的基礎(chǔ)上--幫助研究人員了解了最佳的表面化合物,,利用了一個簡單的方法,,提出了一個合理劃算的提高干細(xì)胞生長速度的方法。研究人員采用了一種創(chuàng)新的,,非常規(guī)的方法提升了聚苯乙烯板中碳?xì)浠衔锖王セ臐舛龋核麄兝昧艘慌_紫外臭氧儀(a UV ozone unit,,張迪編譯),研究人員發(fā)現(xiàn)經(jīng)過2分半鐘的紫外照射后,,培養(yǎng)細(xì)胞就能在無飼養(yǎng)細(xì)胞的條件下快速生長,。(生物谷Bioon.com)
doi:10.1073/pnas.1114854108PMC:
PMID:
Surface-engineered substrates for improved human pluripotent stem cell culture under fully defined conditions
Krishanu Saha, Ying Mei, Colin M. Reisterer, Neena Kenton Pyzocha, Jing Yang, Julien Muffat, Martyn C. Davies, Morgan R. Alexander, Robert Langer, Daniel G. Anderson, and Rudolf Jaenisch
The current gold standard for the culture of human pluripotent stem cells requires the use of a feeder layer of cells. Here, we develop a spatially defined culture system based on UV/ozone radiation modification of typical cell culture plastics to define a favorable surface environment for human pluripotent stem cell culture. Chemical and geometrical optimization of the surfaces enables control of early cell aggregation from fully dissociated cells, as predicted from a numerical model of cell migration, and results in significant increases in cell growth of undifferentiated cells. These chemically defined xeno-free substrates generate more than three times the number of cells than feeder-containing substrates per surface area. Further, reprogramming and typical gene-targeting protocols can be readily performed on these engineered surfaces. These substrates provide an attractive cell culture platform for the production of clinically relevant factor-free reprogrammed cells from patient tissue samples and facilitate the definition of standardized scale-up friendly methods for disease modeling and cell therapeutic applications.