Perfusate, enzymatic solution and method for isolating placental hematopoietic stem cells

Abstract

The invention discloses an irrigating solution, an enzymatic hydrolysate and a method for isolating placenta hematopoietic stem cells. The irrigating solution consists of a penicillin-streptomycin double-antibody, phentolamine, vitamin C and PBS. The enzymatic hydrolysate consists of a basal culture medium, collagenase VIII, pancreatin, 1*heparin sodium, phentolamine, vitamin C and Z-VAD-FMK. The method comprises the following steps: irrigating and cleaning a preprocessed placenta with the irrigating solution; irrigating the enzymatic hydrolysate from placental artery until the enzymatic hydrolysate flows out of placental vein, ligating placental artery and vein and standing by; loosening the placental artery and vein and irrigating the irrigating solution from the artery, collecting all liquid from the vein, centrifuging and reserving cell sediments, and recovering and purifying mononuclear cells containing the hematopoietic stem cells sequentially by virtue of a hydroxyethyl starch method and a red blood cell lysis method; re-suspending and obtaining the placenta hematopoietic stem cells. The irrigating solution, the enzymatic hydrolysate and the method disclosed by the invention can significantly improve separation and extraction efficiencies, and a great amount of cells are obtained; the hematopoietic stem cells are high in content, high in survival rate and quite low in cell pollution rate.

Description

technical field

[0001] The invention relates to the field of stem cells, in particular to a perfusate, an enzymolysis solution and a method for isolating placental hematopoietic stem cells. Background technique

[0002] Hemopoietic Stem cells (HSCs) are a group of primitive hematopoietic cells that exist in hematopoietic tissues. They are not tissue-fixed cells and can exist in hematopoietic tissues and blood. Hematopoietic stem cells are the origin of all hematopoietic and immune cells.

[0003] With the deepening of research, HSC transplantation has become an effective means of treating many malignant hematological diseases and other tumors. At present, the main source of HSC is bone marrow, peripheral blood and cord blood. The proliferation and differentiation ability of bone marrow and peripheral blood HSC is reduced, and it is easily affected by the health status of the donor. However, the number of cord blood HSC is limited. Increasing demand for clinical application...

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