Separation of hematopoietic stem cells from human peripheral blood through modified polyurethane foaming membranes

Abstract

Cell separation from peripheral blood was investigated using polyurethane (PU) foam membranes having 5.2 mum pore size and coated with Pluronic F127 or hyaluronic acid. The permeation ratio of hematopoietic stem cells (CD34(+) cells) and lymphocytes through the membranes was lower than for red blood cells and platelets. Adhered cells were detached from membrane surfaces using human serum albumin (HSA) solution after permeation of blood through the membranes, allowing isolation of CD34(+) cells in the permeate (recovery) solution. High-yield isolation of CD34(+) cells was achieved using Pluronic-coated membranes. This was because the Pluronic coating dissolved into the recovery solution at 4 degrees C, releasing adhered cells from the surfaces of the membranes during permeation of HSA solution through these membranes. Dextran and/or bovine serum albumin solutions were also evaluated for use as recovery solutions after blood permeation. A high recovery ratio of CD34(+) cells was achieved at 4 degrees C in a process using 20% dextran solution through PU membranes having carboxylic acid groups.

Separation of hematopoietic stem and progenitor cells from human peripheral blood through polyurethane foaming membranes modified with several amino acids

“…The blood (feed solution) was subsequently injected into the Syringe 2 described in a previous study. [7][8][9] Two syringes (Syringe 1 and Syringe 2) were attached by head to head. Water was injected into Syringe 1 using perister pump (MP-3, Tokyo Rikakiki Co.) at the speed of 0.3 mL/ min, and therefore, head of Syringe 1 pushed back Syringe 2 containing blood subsequently.…”

“…Finally, the sample was analyzed using the flow cytometry, and the number of red blood cells, platelets, and lymphocytes was counted. [7][8][9] The number of T and B cells in the feed (peripheral blood) and permeate solutions was analyzed from surface markers of CD3 and CD19, respectively. The number of helper T cells and suppressor T cells in the feed (peripheral blood) and permeate solutions was analyzed from surface markers of CD4 and CD8, respectively.…”

“…In this study, we prepared not only the PUACOOH membranes used in our previous studies, [7][8][9] but also different types of PUACOOH membranes modified with several amino acids, and conducted further experiments on the separation of CD34 þ cells using these modified membranes. Furthermore, we used a variety of experimental conditions in the permeation and recovery processes, and investigated the effect of permeation volume, type, and concentration of recovery solution on the detachment of CD34 þ cells (i.e., recovery ratio of CD34 þ cells) during the recovery process at 25 C.…”

“…23 Therefore, the transplantation of CD34 þ cells is required in the therapy of patients with acute myeloid leukemia, chronic myeloid leukemia, myelodysplastic syndromes, and systemic mastocytosis. 24 In previous investigations, [7][8][9] cell separation from peripheral blood was investigated using surface-modified polyurethane (PU) membranes with a pore size of 5 lm, carrying different functional groups. CD34 þ cells adhered more strongly to the membranes than red blood cells, platelets, T cells, or B cells.…”

“…[6][7][8][9] Efficient cell separation is an important issue for the successful isolation and purification of blood cells, stem cells, and specific tissue cells. Centrifugation, 10,11 affinity column chromatography, 12,13 fluorescence-activated cell sorting (FACS), 14,15 magnetic cell selection (MACS), 16,17 and membrane filtration [7][8][9][18][19][20] are techniques typically employed for cell separation. Highly purified cellular preparations are obtained using FACS or MACS in conjunction with a fluorescently labeled antibody as the cell-surface marker, whereas the centrifugal separation of cells is commonly used to isolate platelets, leukocytes, mononuclear cells, red blood cells, and non-blood cells.…”