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MC3T3 细胞系适应杜尔贝科改良伊格尔培养基。

Adaptation of MC3T3 cell line to Dulbecco's Modified Eagle's medium.

机构信息

Nervous System Stem Cells Research Center, Semnan University of Medical Sciences, Semnan, Iran; Department of Tissue Engineering and Applied Cell Sciences, School of Medicine, Semnan University of Medical Sciences, Semnan, Iran.

Nervous System Stem Cells Research Center, Semnan University of Medical Sciences, Semnan, Iran; Department of Tissue Engineering and Applied Cell Sciences, School of Medicine, Semnan University of Medical Sciences, Semnan, Iran.

出版信息

Tissue Cell. 2020 Jun;64:101341. doi: 10.1016/j.tice.2020.101341. Epub 2020 Jan 23.

Abstract

Seeding cells directly into a new medium subjects the cells to stress due to certain differences in medium formulation. As a result, it seems necessary for cells to be adapted to a new medium, in order to save the properties of the cells and to achieve reliable results from the tests. The MC3T3 osteoblastic cell line is recommended to be cultured in Alpha Minimum Essential Medium (α-MEM). However, Dulbecco's Modified Eagle's medium (DMEM) is widely used for its culture. Therefore, in the present paper, two sequential methodologies were applied to adapt the MC3T3 cells to DMEM. In sequential adaptation 1, 10 vol.% DMEM was added to the original medium every day, while in sequential adaptation 2, the old medium was changed to a new medium having 20 vol.% higher DMEM content after each passage. Cells were monitored and compared to direct cell adaptation, while they were growing. The results showed that in the direct cell adaptation, increase in the number of cells was very slow. In contrast, the two sequential adaptation processes were more efficient where sequential adaptation 2 resulted in a higher number of cells in fewer days; 88 % greater than sequential adaptation 1 when it was believed that the cells were adapted. Furthermore, the statistical analysis was conducted by stepwise regression analysis and mathematical models were provided, which can predict the number of cells by day of culture.

摘要

将细胞直接播种到新的培养基中会导致细胞因培养基配方的某些差异而产生应激。因此,似乎有必要使细胞适应新的培养基,以保存细胞的特性,并从测试中获得可靠的结果。MC3T3 成骨细胞系被推荐在α-最低必需培养基(α-MEM)中培养。然而,DMEM 广泛用于其培养。因此,在本研究中,采用了两种连续的方法使 MC3T3 细胞适应 DMEM。在连续适应 1 中,每天向原始培养基中添加 10 体积%的 DMEM,而在连续适应 2 中,每次传代后,将旧培养基更换为 DMEM 含量高 20 体积%的新培养基。在细胞生长过程中对其进行监测并与直接细胞适应进行比较。结果表明,在直接细胞适应中,细胞数量的增加非常缓慢。相比之下,两个连续的适应过程效率更高,连续适应 2 导致在更少的天数内获得更多的细胞;当认为细胞已经适应时,比连续适应 1 多 88%。此外,还进行了逐步回归分析的统计分析,并提供了数学模型,可以通过培养天数预测细胞数量。

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