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.

生物医学科学技术领域对特定细胞群具有特异性亲和力的材料有着强烈的需求。本研究的目的是对淋巴细胞亚群具有特异性亲和力的新型材料的分子设计。由于细胞质膜上表达的酸性官能团的数量和种类不同，我们特别关注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成分，以期通过离子力与 B 细胞和 T 细胞发生特异性相互作用。 B细胞和T细胞是淋巴细胞的两个主要亚群，它们的分离在基础和临床免疫学领域都极其重要。通过对一系列具有聚胺微结构域的新型多相材料，聚HEMA-聚胺与细胞相互作用的系统研究研究发现，接枝共聚物 (HA) 可选择性地从大鼠肠系膜淋巴结淋巴细胞的异质混合物中吸附 B 细胞。对于B细胞和T细胞的分离，对于Mn为3750-6600的聚胺接枝物，HA共聚物中光学聚胺含量为13wt％（HA13），对于Mn为11000的聚胺接枝物，HA共聚物中的光学聚胺含量为7wt％（HA7）。选择性降低对于具有 Mn 小于 1000 的聚胺接枝物的 HA 共聚物，与 HEMA 的无规共聚物一样， N,N'-二乙基-n-(4-乙烯基苯乙基)乙二胺。在分离B细胞和T细胞方面，使用含有13wt%多胺接枝物且Mn为3600-6600的HA接枝共聚物获得了最优异的结果。通过使用 HA13(3750)，获得纯度超过 92% 的 T 细胞群，柱流出物收率为 95%。此外，通过温和的移液，从柱中以 85% 的产率回收了纯度为 90% 的 B 细胞群。因此，我们的结果证明了细胞吸附色谱作为细胞分离的主要工具的一个有前途的特征。