Brodsky Sergey V, Merks Roeland M H, Mendelev Natalia, Goo Cara, Chen Jun
Department of Pathology, New York Medical College, Valhalla, NY 10595, USA.
Diabetes Res Clin Pract. 2007 Jun;76(3):463-7. doi: 10.1016/j.diabres.2006.10.003. Epub 2006 Nov 3.
Studies of cell-matrix, cell-cell interaction or angiogenesis on different matrices require simultaneous comparison of read-out parameters from differently treated companion cells. The culture conditions (cell number, temperature and volume of culture medium) in different chambers are not completely equalized using conventional methods. It has been reported that cells growing in different environmental conditions may exhibit different proliferation patterns [P. Tracqui, J.W. Liu, O. Collin, J. Clement-Lacroix, E. Planus, Global analysis of endothelial cell line proliferation patterns based on nutrient-depletion models: implications for a standardization of cell proliferation assays, Cell Proliferat. 38(June (3)) (2005) 119-135]. Herein we describe an innovative chamber, which could resolve this problem by significantly improving the standardization of experimental conditions. The chamber was manufactured from a standard cell culture well by its division with a septum into two sections. We utilized the chamber and recently developed topological analysis to examine the effects of glycated matrices on the capillary-like network formation by endothelial cells. Glycated Collagen I resulted in dose-dependent changes to all measured topological characteristics of the capillary-like network, such as the number of branching points, number of meshes and total capillary length. These differences were observed only in neighbored compartments coated with different matrices, but not in the compartments coated with the same matrix. The novel chamber brings an opportunity for better standardization of experimental conditions and simultaneous observation of different experimental groups, reducing the possible effect of any systematic error.
对不同基质上的细胞 - 基质、细胞 - 细胞相互作用或血管生成的研究需要同时比较不同处理的伴随细胞的读出参数。使用传统方法无法完全均衡不同培养室中的培养条件（细胞数量、温度和培养基体积）。据报道，在不同环境条件下生长的细胞可能表现出不同的增殖模式[P. Tracqui，J.W. Liu，O. Collin，J. Clement - Lacroix，E. Planus，基于营养耗尽模型的内皮细胞系增殖模式的全局分析：对细胞增殖测定标准化的影响，《细胞增殖》。38（6月（3））（2005年）119 - 135]。在此，我们描述了一种创新的培养室，它可以通过显著提高实验条件的标准化来解决这个问题。该培养室由标准细胞培养孔制成，通过隔膜将其分成两部分。我们利用该培养室和最近开发的拓扑分析来研究糖化基质对内皮细胞形成毛细血管样网络的影响。糖化I型胶原蛋白导致毛细血管样网络的所有测量拓扑特征出现剂量依赖性变化，如分支点数、网眼数和总毛细血管长度。这些差异仅在涂有不同基质的相邻隔室中观察到，而在涂有相同基质的隔室中未观察到。这种新型培养室为更好地标准化实验条件和同时观察不同实验组带来了机会，减少了任何系统误差的可能影响。
