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

形成血液和免疫细胞的干细胞被称为造血干细胞。造血干、祖细胞带有CD34细胞表面标志。这些细胞被认为负责重建造血系统。因此，CD34细胞的移植在治疗急性髓系白血病、骨髓增生异常综合征、慢性髓系白血病和系统性肥大细胞增多症患者中是必不可少的。采用(A)未修饰的聚氨酯(PU)，(B)表面修饰的氨基、磺酸、RGDS和Redv多肽的PU，(C)Pluronic固定化的PU，(D)细胞黏附分子(胶原、纤维连接蛋白、层粘连蛋白和Vitronectin)固定的PU和PU-COOH膜，研究了从外周血和骨髓中分离干细胞的方法。无论使用何种类型的PU膜，造血干细胞(CD34细胞)通过这些膜的透过率都是血细胞中最低的，而红细胞和血小板通过PU膜的通透率更高。这是因为造血干细胞倾向于最强烈地附着在膜表面，因为它们的表面高表达细胞黏附分子。通过蛋白质定量试剂盒(微量BCA测量)、XPS和ATR-FT-IR分析了膜上引入的纳米片段的数量。进一步研究了通过表面改性尼龙网状膜的过滤方法从小鼠骨髓中分离骨髓间充质干细胞，并在一定程度上实现了骨髓间充质干细胞的纯化。

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.