Method for amplifying candidate stem cell
A technology of hematopoietic stem cells and nuclear cells, which can be applied to blood/immune system cells, animal cells, vertebrate cells, etc., and can solve the problems of anti-leukemia and graft-versus-host effect changes, inability to meet clinical use requirements, and reduced proliferation ability. , to reduce the differentiation of hematopoietic stem cells, meet the requirements of clinical use, and promote the generation of stromal cells
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Embodiment 1
[0033] A method of expanding hematopoietic stem cells,
[0034] 1) Isolation of mononuclear cells:
[0035]Acquisition of nucleated cells from human umbilical cord blood: add methylcellulose solution with a weight concentration of 0.125% to human umbilical cord blood at a volume ratio of 4:1, let stand at room temperature, and settle red blood cells; take the supernatant at 1500rpm / min Centrifuge, and the sediment is nucleated cells.
[0036] Density gradient centrifugation, add Ficoll lymphocyte separation medium with a density of 1.077 / ml to the nucleated cells at a volume ratio of 1:1, 1500rpm / mim, 30min, absorb the mononuclear cells in the middle layer; it contains CD34 + The number of cells is 0.9% of the total number of cells; the cell density is adjusted after washing with PBS (1500 rpm / min, 5 min).
[0037] 2) Preparation of microencapsulated mononuclear cells:
[0038] A. Sodium alginate with a weight content of 31.2% of β-L-guluronic acid and a weight content of 6...
Embodiment 2
[0051] A method of expanding hematopoietic stem cells,
[0052] 1) Isolation of mononuclear cells:
[0053] Density gradient centrifugation, add Ficoll lymphocyte separation medium with a density of 1.077 / ml to the human bone marrow fluid at a volume ratio of 1.5:1, 1500rpm / mim, 30min, absorb the mononuclear cells in the middle layer; it contains CD34 + The number of cells is 1.9% of the total number of cells; the cell density is adjusted after washing with PBS (1500rpm / min, 5min);
[0054] 2) Preparation of microencapsulated mononuclear cells:
[0055] A. Sodium alginate with a weight content of 37% of β-L-guluronic acid and a weight content of 63% of α-D-mannuronic acid was prepared into 1.0% w / v sodium alginate solution I;
[0056] Sodium alginate containing 62% by weight of β-L-guluronic acid and 38% by weight of α-D-mannuronic acid was formulated into 2.5% w / v sodium alginate solution II;
[0057] Mix sodium alginate solution I and sodium alginate solution II at a volu...
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