Nanoporous Membranes for cellular microarrays and in vitro assays

用于细胞微阵列和体外测定的纳米多孔膜

基本信息

批准号：
8200228
负责人：
THOMAS R GABORSKI
金额：
$ 18.47万
依托单位：
SIMPORE, INC.
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-09-01 至 2012-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8200228
关键词：
Address Adherent Culture Apical Area Basic Science Behavioral Biological Assay Caliber Cell Culture Techniques Cell Survival Cells Cellular Membrane Characteristics Coculture Techniques Custom DNA Microarray Chip Development Devices Diffusion Drug Formulations Endothelial Cells Environment Equipment Glass Goals Growth Health Home environment Human Image Image Analysis Island Licensing Life Measurement Measures Membrane Miniaturization Molecular Paper Pattern Permeability Pharmaceutical Preparations Phase Population Porosity Preclinical Drug Evaluation Protein Microchips Publishing Reagent Research Research Personnel Resistance Robotics Screening procedure Series Side Silicon Solutions Sorting - Cell Movement Standardization Stem cells Stimulus Supporting Cell Surface Suspension Culture Suspension substance Suspensions System Techniques Technology Testing Thick Thinness Time Tissue Engineering Tissues Umbilical vein Universities Variant Work cancer cell cell growth cost culture plates cytotoxicity cytotoxicity test density drug development drug discovery fluorescence imaging high throughput screening in vitro Assay miniaturize monolayer nanocrystal nanometer novel prototype research and development research study response stem cell differentiation tool

项目摘要

DESCRIPTION (provided by applicant): Advances in protein and DNA microarrays have enabled dramatic increases in throughput and equipment standardization has made these techniques more commonplace. High density, high throughput microarrays reduce the cost of research and development in drug discovery and basic science by decreasing reagent volumes and increasing the number of experiments per plate. Missing from this miniaturization, however, are cell culture microarrays. Existing low well count cell culture plates require greater volumes of precious drug formulations for permeability assays and more plates are required to complete a series of experiments. These same factors increase the cost of parallelized cellular experimentation in basic science such as screening stem cell culture differentiation conditions. In this proposal we will test the feasibility of using a new class of ultrathin nanoporous membrane to enable miniaturization of cell culture screening for high throughout drug permeability and co-culture studies. At the limit we will enable single cell screening to study phenotypic and behavioral variations in cell populations in response to stimuli, drug treatments or co-culture environments. In the first Aim of this work, we will fabricate microarray-scale cell culture arrays using porous nanocrystalline silicon (pnc-Si). We will confirm these devices and size format promote healthy growth of primary human umbilical vein endothelial cells by comparing cytotoxicity and growth curve measurements against larger conventional cell inserts. To test feasibility as a high throughout platform for single cell and co-culture screening, we will develop a microarray of wells on pnc-Si. Our approach is novel because we will be the first to offer a membranesupported microarray that enables study variations in populations of cancer cells, stem cells as well as primary cell response to drug treatment in a co-culture environment. In Phase II we will focus on drug screening and stem cell differentiation with the goal of developing an automated cell dispensing and fluorescent image analysis system. In both cases we will also pursue enlarged microarrays (>100 microns) with degradable membrane supports, which will permit the growth a small islands of stratified tissue. Successful completion of Phase I will enable the launch of a live imaging research tool for small-scale cell co-culture. Within 6 months of completing Phase II, we will introduce a 384-window microarray system with >10x5 wells.

描述（由申请人提供）：蛋白质和DNA微阵列的进步使吞吐量和设备标准化的急剧增加使这些技术更加普遍。高密度，高吞吐量微阵列通过减少试剂量并增加每个板的实验数量，从而降低了药物发现和基础科学中的研发成本。然而，这种微型化的缺少是细胞培养的微阵列。现有的低井计数细胞培养板需要大量的宝贵药物制剂来进行渗透性测定，并且需要更多的板才能完成一系列实验。这些相同的因素增加了基础科学中平行细胞实验的成本，例如筛选干细胞培养分化条件。在此提案中，我们将测试使用新型的超薄纳米多孔膜的可行性，以使整个药物渗透性和共培养研究中的细胞培养筛查能够微型化。在限制下，我们将使单细胞筛选能够研究细胞种群的表型和行为变化，以响应刺激，药物治疗或共培养环境。在这项工作的第一个目的中，我们将使用多孔纳米晶硅（PNC-SI）制造微阵列规模的细胞培养阵列。我们将通过比较细胞毒性和生长曲线测量值与较大的常规细胞插入物进行比较，确认这些设备和尺寸格式促进原代人脐静脉内皮细胞的健康生长。为了测试可行性，作为单元和共培养筛查的整个平台的高度，我们将在PNC-SI上开发Wells的微阵列。我们的方法是新颖的，因为我们将是第一个提供膜支持的微阵列的人，可以研究在共培养环境中对癌细胞，干细胞以及对药物治疗的原发性细胞反应的研究变化。在第二阶段，我们将专注于药物筛查和干细胞分化，以开发自动细胞分配和荧光图像分析系统。在这两种情况下，我们还将使用可降解的膜支撑进行扩大的微阵列（> 100微米），这将允许生长一个小岛分层组织。成功完成一阶段将启动用于小型细胞共文化的实时成像研究工具。在完成II期的6个月内，我们将引入一个384窗口的微阵列系统，其井中> 10x5井。