CELL PARTITIONING WITH TWO PHASE AQUEOUS POLYMERS

用两相水性聚合物进行细胞划分

• 批准号: 3337662
• 负责人: GEOFFREY V SEAMAN
• 金额: $ 10.97万
• 依托单位: OREGON HEALTH & SCIENCE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1979
• 资助国家: 美国
• 起止时间: 1979-07-01 至 1986-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3337662
• 关键词: analytical method; cell sorting; chemical models; clinical biomedical equipment; dextrans; erythrocytes; hereditary spherocytosis; human subject; human tissue; membrane activity; membrane potentials; multiple sclerosis; paroxysmal nocturnal hemoglobinuria; polymers; thermodynamics; water solution

Partitioning of cells and membranes in tow-polymer aqueous phase systems by means of counter current distribution (CCD) has been shown empirically to be a sensitive, versatile technique for the characterization and fractionation of cell subpopulations on the basis of surface or membrane properties. It is propsed to continue evaluation and development of a thermodynamic model of cell partitioning in such systems, since the partition theory developed to date predicts much higher resolution separations than are observed. Different phase systems will be examined containing variety of anions, especially phosphate and sulfate. The systems will be characterized in terms of polymer content, salt composition, density and viscosity of the phases, interfacial tensions, and also electrostatic and surface potentials of the phases. The effects of poymer molecular weight and polymer chemical composition will be examined by using PEG-dextran and ficoll-dextran systems with polymers of various molecular weights. Other determinants of partitioning will be examined including measurement of three phase contact angles on single cells adsorbed at the phase boundary, cell-phase droplet interactions and influence of fluid shear and convection in determining partition. Well characterized model particles of defined sizes and surface properties such as monodisperse polystyrene latices will be used to evaluate some of the predicted primary determinants of particle phase partitioning. The experimental and system variables of the CCD method will be evaluated and new techniques explored for the separation of cell populations which differ only slightly in surface properties. The knowledge gained from the basic studies will be used to examine the partitioning behavior of erythrocytes from patients with multiple sclerosis (MS), hereditary spherocytosis and paroxysmal nocturnal hemoglobinuria (PNH). In the case of MS is is hoped to develop a test for the diagnosis of the disease which in addition could yield new mechanistic information on MS, and with PNH preliminary CCD studies have enabled us to nondestructively isolate abnormal RBC subpopulations.

双聚合物水相体系中细胞和膜的分离 逆流分布方法(CCD)已经被经验证明是 是一种敏感的、多功能的技术，用于表征和 基于表面或膜的细胞亚群的分级 属性。建议继续评估和开发一种 这类系统中细胞划分的热力学模型，因为 到目前为止发展起来的分割理论预测了更高的分辨率 分离比观察到的要多。不同的阶段系统将被检查 含有各种阴离子的，尤指磷酸盐和硫酸盐。这个 系统将根据聚合物含量、盐 相的组成、密度和粘度、界面张力以及 以及相的静电势和表面电势。的影响 将检查聚合物的相对分子质量和聚合物的化学组成。 通过将聚乙二醇-葡聚糖和菲科尔-葡聚糖体系与不同种类的聚合物一起使用 分子量。我们还将研究分区的其他决定因素 包括单电池三相接触角的测量 在相边界上的吸附，细胞-相液滴的相互作用和 流体剪切和对流对确定分配的影响。井 表征定义大小和表面属性的模型粒子，如 作为单分散聚苯乙烯胶乳将被用来评估一些 预测了颗粒相分配的主要决定因素。这个 将对CCD方法的实验和系统变量进行评估和 探索分离不同细胞群的新技术 仅在表面属性上略有变化。从基础知识中获得的知识 研究将用于检查红细胞的分配行为 来自多发性硬化症(MS)、遗传性球形红细胞增多症和 阵发性睡眠性血红蛋白尿(PNH)在MS的情况下，希望是 开发一种诊断这种疾病的测试，此外，它还可以 在MS上产生新的机理信息，并与PNH初步CCD 研究使我们能够无损地分离异常的红细胞 亚群。