Perfusate, enzymatic solution and method for separating 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 I, collagenase II, heparin sodium, phentolamine, vitamin C and Z-VAD-FMK. The method comprises the following steps: cleaning blood inside a placenta with the irrigating solution; irrigating the enzymatic hydrolysate into the placenta and enzymatically hydrolyzing at room temperature; collecting the enzymatic hydrolysate, centrifuging and reserving cell sediments; re-suspending the cell sediments by virtue of the PBS, adding a hydroxyethyl starch solution, standing by, reserving and centrifuging upper liquid and discarding supernatant; re-suspending the cell sediments by virtue of a red blood cell lysis solution, promoting lysis to residual red cells, centrifuging and discarding supernatant; and re-suspending the cell sediments by virtue of a culture medium, centrifuging and discarding supernatant, so that the placenta hematopoietic stem cells are obtained. 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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