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Preparation of cell separation membranes and stem cell separation for regenerative medicine from bone marrow and blood.

骨髓和血液再生医学细胞分离膜的制备和干细胞分离。

基本信息

批准号：
17560691
负责人：
HIGUCHI Akon
金额：
$ 2.37万
依托单位：
National Research Institute for Child Health and Development (2007)Seikei University (2005-2006)
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2005
资助国家：
日本
起止时间：
2005 至 2007
项目状态：
已结题

项目摘要

The stem cells that form blood and immune cells are known as hematopoietic stem cells. Hematopoietic stem and progenitor cells bear the CD34 cell surface marker. These cells are thought to be responsible for the reconstitution of hematopoiesis. Therefore, the transplantation of CD34 cells is essential in the therapy of patients with acute myeloid leukemia, myelodysplastic syndromes, chronic myeloid leukemia, and systemic mastocytosis.Stem cell separation from peripheral blood and bone marrow was investigated using (a) unmodified polyurethane (PU), (b) surface-modified PU having amino group, sulfonic acid, RGDS peptide, and REDV peptide (c) Pluronic-immobilized PU, (d) cell adhesive molecules (collagen, fibronectin, laminin and vitronectin)-immobilized PU, and PU-COOH membranes. The permeation ratio of hematopoietic stem cells (CD34 cells) through these membranes was the lowest among blood cells regardless of the type of PU membrane used, while red blood cells and platelets permeated more freely through the PU membranes. This was because hematopoietic stem cells tend to adhere the strongest to membrane surfaces, due to their high expression of cell-adhesion molecules on their surfaces. Amount of nano-segments introduced on the membranes was analyzed from protein quantification kit (micro BCA measurement), XPS, and ATR-FT-IR.We further investigate the separation of mesenchymal stem cells from mice bone marrow by the filtration method through surface-modified nylon mesh membranes, and purification of mesenchymal stem cells was achieved in some extent.

形成血液和免疫细胞的干细胞被称为造血干细胞。造血干细胞和祖细胞具有CD 34细胞表面标记。这些细胞被认为是负责造血重建。使用（a）未改性的聚氨酯（PU），（B）具有氨基、磺酸、RGDS肽和REDV肽的表面改性的PU，（c）Pluronic固定的PU，（d）细胞粘附分子（胶原、纤连蛋白、层粘连蛋白和玻连蛋白）固定的PU和PU-COOH膜。无论使用哪种类型的PU膜，造血干细胞（CD 34细胞）通过这些膜的渗透率在血细胞中是最低的，而红细胞和血小板则更自由地通过PU膜渗透。这是因为造血干细胞倾向于最强地粘附到膜表面，这是由于其表面上的细胞粘附分子的高表达。通过蛋白质定量试剂盒（microBCA measurement）、XPS、ATR-FT-IR分析膜上纳米链段的引入量，进一步研究了表面改性尼龙网膜过滤法分离小鼠骨髓间充质干细胞的效果，并在一定程度上实现了间充质干细胞的纯化。