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

• 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
• 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

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(申请人提供)：蛋白质和DNA微阵列的进步使产量大幅增加，设备标准化使这些技术变得更加常见。高密度、高通量的微阵列通过减少试剂体积和增加每个平板的实验数量来降低药物发现和基础科学的研究和开发成本。然而，在这种小型化过程中，缺失了细胞培养微阵列。现有的低孔数细胞培养板需要更多的珍贵药物配方来进行渗透性分析，并且需要更多的板来完成一系列实验。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。在这项提议中，我们将测试一种新型超薄纳米多孔膜的可行性，以实现细胞培养的微型化，筛选高通量药物渗透和共培养研究。在这一限制下，我们将使单细胞筛选能够研究细胞群体对刺激、药物治疗或共培养环境的表型和行为变化。在这项工作的第一个目标，我们将制造微阵列规模的细胞培养阵列使用多孔纳晶硅(PNC-Si)。我们将通过比较细胞毒性和生长曲线测量与较大的常规细胞插入物来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。为了测试作为单细胞和共培养筛选的高通量平台的可行性，我们将在PNC-Si上开发微阵列。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，可以在共培养环境中研究癌细胞、干细胞以及原代细胞对药物治疗的反应的变化。在第二阶段，我们将专注于药物筛选和干细胞分化，目标是开发一种自动化细胞分配和荧光图像分析系统。在这两种情况下，我们还将寻求具有可降解膜支撑物的扩大的微阵列(&gt；100微米)，这将允许层状组织的小岛生长。第一阶段的成功完成将使小规模细胞共培养的活体成像研究工具得以推出。在完成第二阶段的6个月内，我们将推出一个拥有10x5个井的384个窗口的微阵列系统。