权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Separation of lymphocyte subpopulations by novel polymeric adsorbent with controlled micro-structure

新型可控微结构聚合物吸附剂分离淋巴细胞亚群

基本信息

批准号：
61890010
负责人：
TSURUTA Teiji
金额：
$ 3.71万
依托单位：
Science University of Tokyo
依托单位国家：
日本
项目类别：
Grant-in-Aid for Developmental Scientific Research
财政年份：
1986
资助国家：
日本
起止时间：
1986 至 1987
项目状态：
已结题

项目摘要

There is a stron demand for the materials which have the specific affinity to the particular cell population in the field or biomedical science and technology. The objective of this study is the molecular design of novel materials with specific affinity to lymphocyte subpopulations. We gave special attention to the diiferencial ionic character of cell surfaces owing to the differ- ences in the number and species of acidic functional groups expressed on their plasma membrane. Thus, basic amino groups were chosen as the main component of our designing polymeric materials to expect the specific interaction with B and T cells through ionic force. B and T cells are the two major subpopulations of lymphocytes, and their saparation is extremely important in the field of fundamental as well as clinical immunology.Through the systematic study on cellular interaction with a series of new multiphase material with polyuamine microdomains, polyHEMA-polyamine graft copolymer (HA) was found to adsorb B cells selectively out of a heterogeneous mixture of rat mesenteric lymphnode lymphocytes. As for the separation of B and T cells, the optical polyamine content in HA copolymer was 13wt% (HA13) for the polyamine grafts with Mn of 3750-6600 and is 7wt% (HA7) for that with Mn of 11000. Selectivity was lowered for HA copolymer having polyamine grafts with Mn of less than 1000 as sell as for ran- dom copolymer of HEMA with N,N'-diethyl-n-(4-vinylphenethyl)ethylenediamine.The most excellent result in terms of separation of B and T cells was obtained with HA graft copolymer with 13wt% of pollyamine grafts with Mn of 3600-6600. By using HA13(3750), T cell popu- lation with more than 92% purity was obtained in 95% yield as the column effluent. Also, by gentle pipetting, B cell population with 90% purity was recovered in 85% yield from the column. Consequently, our results demonstrated a promising feature of cell adsorption chromatography as a major tool for cell separation.

在生物医学科学和技术领域中，对与特定细胞群体具有特异性亲和性的材料有着强烈的需求。本研究的目的是设计对淋巴细胞亚群具有特异性亲和力的新型材料。由于细胞质膜上酸性官能团的数量和种类不同，我们特别注意细胞表面的不同离子特征。因此，选择碱性氨基作为我们设计的聚合物材料的主要组分，期望通过离子力与B和T细胞特异性相互作用。B细胞和T细胞是淋巴细胞的两个主要亚群，它们的分离在基础免疫学和临床免疫学领域都是非常重要的.通过对一系列新型多胺微区多相材料与细胞相互作用的系统研究，发现多聚HEMA-多胺接枝共聚物（HA）能从大鼠肠系膜淋巴结淋巴细胞的异质混合物中选择性地吸附B细胞.对于B和T细胞的分离，对于Mn为3750-6600的多胺接枝物，HA共聚物中的最佳多胺含量为13wt%（HA 13），对于Mn为11000的多胺接枝物，HA共聚物中的最佳多胺含量为7wt%（HA 7）。具有Mn小于1000的多胺接枝物的HA共聚物的选择性与HEMA与N，N ′-二乙基-N-（4-乙烯基苯乙基）乙二胺的无规共聚物一样降低，在分离B和T细胞方面，具有13wt%的多胺接枝物的HA接枝共聚物（Mn为3600-6600）获得最优异的结果。用HA 13（3750）作为柱流出物，以95%的收率获得纯度大于92%的T细胞群。此外，通过轻轻移液，从柱中以85%的产率回收纯度为90%的B细胞群。因此，我们的研究结果表明，一个有前途的功能，细胞吸附色谱作为一个主要的工具，细胞分离。