• 再生营养物质培养环境下的细胞择选
  • Cellular Selection in the Cell Culture Environment Created by a Regenerative Nutritional Substance
  • 徐鹏,张丽,陶国新.再生营养物质培养环境下的细胞择选[J].中国烧伤创疡杂志,2012,24(1):1~12.
    DOI:10.3969/j.issn.1001-0726.[year].[issue].[sequence]
    中文关键词:  再生物质  细胞择选  共培养  细胞增殖
    英文关键词:Regenerative substance,Cellular selection,Co-culture,Cell proliferation
    基金项目:
    作者单位
    徐鹏 北京荣祥再生医学研究所 
    张丽 北京荣祥再生医学研究所 
    陶国新 北京荣祥再生医学研究所 
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    中文摘要:
          在加入再生营养物质(再生物质)油膏剂缓释物的培养基中,共培养细胞中癌细胞的增殖受到选择性抑制,但是在细胞体外培养实验中,油膏剂难以溶解于培养基中。利用新研制的水溶性再生物质J,对人胚肺二倍体成纤维细胞MRC5(MRC5细胞)和人肺腺癌细胞A549(A549细胞)进行体外共培养,实验结果显示,加入了体积分数为0.015% ~0.02%的J的培养基既能够选择性高效抑制A549细胞增殖,又能维持MRC5细胞存活和增殖。这一体积分数范围的J的培养基代表了一种逆转A549细胞对MRC5细胞竞争优势的营养环境,就像自然加入了该选择一样,共培养环境中的细胞在适应环境过程中也在进行着生存竞争。既往的研究发现,以上两种细胞在适应含有再生物质的培养环境时,线粒体有明显不同的变化,我们进一步对MRC5细胞在再生营养环境中进行传代培养,并测定细胞中与线粒体生成和能量代谢相关的SIRT1蛋白表达的变化,发现在体积分数为0.015% ~0.02%的J的培养基中传代的MRC5细胞SIRT1蛋白表达接近于未加再生物质的对照组,而其他体积分数J的培养基与其他再生物质的培养基中的MRC5细胞SIRT1蛋白表达明显不同于对照组的模式。结合共培养的实验结果,在体积分数为0.015% ~0.02%的J的培养基营养环境中,很可能是MRC5细胞可以适应并能够维持正常能量代谢,但是A549细胞不能适应且能量代谢失调,从而使MRC5细胞在此环境下的细胞择选中胜出。
    英文摘要:
          In the cell culture environment having contents slowly released into the medium from a regenerative nutritional substance (regenerative substance) oil/ointment, the proliferation of cancer cells in a co-culture system is selectively inhibited while the water solubility of regenerative substance remains a challenge in in vitro cell culture. With the application of a newly developed water soluble formula of the regenerative substance J, we conducted a co-culture of the MRC5 lung cell line and the A459 lung cancer cell line. We identified the concentration range of J (0.015%-0.02%) in the culture medium in which the proliferation of the A459 cell line was selectively and efficiently inhibited while the survival and proliferation of the MRC5 cell line was maintained. The culture medium of 0.015%-0.02% J indicates a nutritional environment which reverses the advantage of the A459 cell line in a survival competition with the MRC5 cell line. Since the mitochondria of the A459 and MRC5 cell lines were found to act in different ways when the cells were cultured in the medium having a regenerative substance in a previous study, we conducted a continuous passage culture of the MRC5 cell line and characterized the expression of SIRT1, a biomarker associated with mitochondrial biogenesis and energy metabolism. The results showed that only when cultured in a medium of 0.015%-0.02% J, MRC5 expressed SIRT1 similarly as it did in a normal control medium, and MRC5 cultured in the medium of other concentrations of the regenerative substance expressed SIRT1 in a way different from the normal control. Taking into account the results of the co-culture of A459 and MRC5 cell lines, we propose that in the nutritional environment of a 0.015%-0.02% J medium, the MRC5 cell line wins the survival competition probably by better adaptation and maintaining, in part, normal energy metabolism while the energy metabolism of cancer cells is disturbed.