Time dependent cell growth in biomedical research with General Cell Screening System

G. Pretnar, F. Steindl
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Abstract

Modern and effective biomedical research needs new and effective methods. Use of 96-well micro-titre plates allows the performance of many experiments and samples under the same conditions. The methods used to determine cell numbers in these plates are all end-point methods. These methods allow only one determination per plate, and the cells are lost. Usually, such assays are performed according to concentration dependance. Other parameters and cell growth dynamics remained unrevealed. To reveal cell growth dynamics, multiple assays should be performed. Such an experimental approach would be expensive and time-consuming, so it is rarely performed. To fill this gap, the Institut fu/spl uml/r Angewandte Mikrobiologie (IAM) and SLT Labinstruments developed GCSS (General Cell Screening System). GCSS is a powerful hardware/software system that enables continuous monitoring of cell growth without any treatment or stain. The method is based on a high-resolution turbidity measurement performed directly on the cell culture plate. The system consists of a reader and an eight-channel photometer, a plate with a new form of wells, an Apple Macintosh computer and the GCSS software. We chose a classic bone-marrow colony-count assay, which is a typical assay scored after seven days of incubation and based on one measurement only. GCSS allowed us to seed bone marrow cells in microtitre plates in a medium with different concentrations of haemopoietic growth factors and to perform multiple measurements. We observed bone-marrow cell growth derived from interferon alpha-treated mice and compare the cell growth from placebo-treated mice. The classical bone marrow assay only allowed scientists to confirm the suppressive nature of interferon alpha on bone marrow cells in-vivo. Multiple measurements with GCSS allowed us to collect cell growth data over seven days of incubation which could not be seen in the colony-count assay. The data collected with GCSS could not confirm the suppressive nature of interferon alpha but revealed that it is a bone-marrow cell activator in-vivo. We expect that GCSS will have an important influence in new biomedical research, and in corrections of established assays too.
通用细胞筛选系统在生物医学研究中的时间依赖性细胞生长
现代有效的生物医学研究需要新的有效方法。使用96孔微量滴定板可以在相同条件下进行许多实验和样品。测定这些板中细胞数目的方法都是终点法。这些方法只允许每块板测定一次,而且细胞会丢失。通常,这种测定是根据浓度依赖性进行的。其他参数和细胞生长动力学仍未揭示。为了揭示细胞生长动态,需要进行多次检测。这种实验性的方法既昂贵又耗时,因此很少被采用。为了填补这一空白,研究所/spl uml/r angelandte微生物(IAM)和SLT Labinstruments开发了GCSS(通用细胞筛选系统)。GCSS是一个强大的硬件/软件系统,可以连续监测细胞生长,无需任何处理或染色。该方法基于直接在细胞培养板上进行的高分辨率浊度测量。该系统由一个读取器、一个八通道光度计、一个带有新型孔的平板、一台苹果Macintosh电脑和GCSS软件组成。我们选择了经典的骨髓菌落计数测定法,这是一种典型的测定法,在孵育7天后评分,仅基于一次测量。GCSS使我们能够在含有不同浓度造血生长因子的培养基中将骨髓细胞植入微滴板中,并进行多次测量。我们观察了干扰素处理小鼠的骨髓细胞生长情况,并比较了安慰剂处理小鼠的细胞生长情况。经典的骨髓实验只允许科学家确认干扰素α在体内对骨髓细胞的抑制性质。使用GCSS进行多次测量,使我们能够收集在孵育7天内的细胞生长数据,这在集落计数试验中是看不到的。GCSS收集的数据不能证实干扰素α的抑制性质,但显示它是一种体内骨髓细胞激活剂。我们预计GCSS将在新的生物医学研究中产生重要影响,并在已建立的测定方法的修正中也会产生重要影响。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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