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

在生物医学科学和技术领域中，对特定细胞群体具有特定亲和力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定亲和力的新型材料。由于细胞质膜上表达的酸性官能团的数量和种类不同，我们特别注意了细胞表面的不同离子特性。因此，我们选择碱性氨基作为我们设计的聚合物材料的主要成分，以期通过离子作用力与B细胞和T细胞发生特异的相互作用。B细胞和T细胞是淋巴细胞的两个主要亚群，其分离在基础免疫学和临床免疫学领域具有极其重要的意义。通过对一系列具有多胺微区的新型多相材料与细胞相互作用的系统研究，发现聚HEMA-多胺接枝共聚物(HA)能够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细胞和T细胞的分离，多胺接枝物中的最佳多胺含量分别为13wt%(HA_(13))和7wt%(HA_7)。与HEMA与N、N‘-diethyl-n-(4-vinylphenethyl)ethylenediamine.The的随机共聚物相比，多胺接枝的HA共聚物的选择性降低。在分离B和T细胞方面，以13wt%的多胺接枝的HA共聚物的选择性最好。用HA13(3750)柱分离纯化T细胞，纯度达92%以上，柱流出液产率达95%。此外，通过温和的移液，从柱子中以85%的产率回收了纯度为90%的B细胞群。因此，我们的结果显示了细胞吸附色谱作为细胞分离的主要工具的一个很有前途的特点。

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.