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

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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.