Fluorescence based cell counting in collagen monolayer cultures of primary hepatocytes
Fluorescence based cell counting in collagen monolayer cultures of primary hepatocytes
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DOI：
10.1007/s10616-014-9821-1
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发表时间：
2016-08-01
期刊：
影响因子：
2.2
通讯作者：
Noor, F.
中科院分区：
文献类型：
--
作者：
Priesnitz, C.;Sperber, S.;Noor, F.
细胞数量的准确测定对于定量描述生物过程是必不可少的。这些变化应与可测量的参考相关，例如，在细胞培养的情况下，活细胞数是一个非常有价值的参考参数。细胞数量/活力测量的间接方法可能具有高达10%的标准偏差。这可能导致在“组学”数据的分析以及时间过程研究中出现不期望的大偏差。这样的数据应该优选地被标准化为在给定时间的确切活细胞数，以允许对生物过程的有意义的解释和理解。细胞数量的手动计数是非常费力的，并且在某些实验设置中是不可能的。因此，我们开发了一种简单可靠的基于荧光的方法，其准确度为95- 98%，用于原位测定活细胞数。我们优化了原代大鼠肝细胞的接种细胞密度，以获得最佳细胞粘附。这将有助于有效地使用可用性通常有限的原电池。该方法将是非常有用的“组学”技术的应用，特别是代谢物组学分析，其中代谢物的吸收/生产的具体速率可以可靠地计算。
Accurate determination of cell number is essential for the quantitative description of biological processes. The changes should be related to a measurable reference e.g. in the case of cell culture, the viable cell number is a very valuable reference parameter. Indirect methods of cell number/viability measurements may have up to 10 % standard deviation. This can lead to undesirable large deviations in the analysis of "-omics" data as well as time course studies. Such data should be preferably normalized to the exact viable cell number at a given time to allow meaningful interpretation and understanding of the biological processes. Manual counting of cell number is very laborious and not possible in certain experimental setups. We therefore, developed a simple and reliable fluorescence based method with an accuracy of 95-98 % for the determination of the viable cell number in situ. We optimized the seeding cell densities for primary rat hepatocytes for optimal cell adhesion. This will help in efficient use of primary cells which are usually limited in availability. The method will be very useful in the application of "-omics" techniques, especially metabolome analysis where the specific rates of uptake/production of metabolites can be reliably calculated.