Method for separating and purifying stem cells from human umbilical cord blood

Abstract

The invention relates to a method for separating and purifying stem cells from human umbilical cord blood. The method comprises the following steps: adopting a kit for separating and purifying human umbilical cord blood enchylema, thereby acquiring a mononuclear cell sample, wherein the kit comprises a reagent A and a reagent B, the reagent A is an erythrocyte precipitator and is composed of 6% hydroxyethyl starch solution and the reagent B is a cell separating medium, is prepared from saccharosan 400 and cardiografin and is a sterile aqueous solution; adopting a stem cell culture medium for culturing, multiplying and purifying the stem cells, wherein the stem cell culture medium is prepared by mixing DMEM/F12 stem cell culture medium and optimized additives. According to the invention, the umbilical cord blood stem cell in vitro culture is utilized and the components are added into the cell culture medium, so that the multiplication advantage of the mesenchymal stem cells is promoted, the growth of other cells is expelled and the cells are naturally purified; the recovery of the monocytes is more than or equal to 95%; the survival rate of the stem cells is more than or equal to 98%; the quantity of the mesenchymal stem cells is more than or equal to 5*10<7>; the quality requirement of clinic treatment is met.

Description

technical field [0001] The invention relates to the technical field of medical experiments, in particular to a method for separating and purifying mesenchymal stem cells from human umbilical cord blood, using the principle of gradient centrifugation to quickly separate large-capacity umbilical cord blood to prepare mononuclear cells, and on this basis, using cell culture Sorting technology prepares stem cells for clinical and laboratory needs. Background technique [0002] With the continuous deepening of mononuclear cell research and clinical application of monocytes, hematopoietic stem cell transplantation has become a radical cure for malignant blood diseases (leukemia, malignant lymphoma, multiple myeloma), aplastic anemia, and certain genetic diseases. effective means. In recent years, stem cell transplantation has been gradually applied in the clinical treatment of heart disease, nervous system injury, tissue and organ repair, diabetes, and vascular disease. In addit...

AI Technical Summary

Problems solved by technology

[0005] At present, the methods for separating and extracting hematopoietic stem cells from umbilical cord blood mainly include flow cytometry, which is expensive in equipment, high in clinical application costs, and inconvenient to popularize; culture expansion method, which has a high pollution rate and takes a long time, is not suitable for clinical use. ; Immunomagnetic bead method, in which the cells are labeled, the screening range is small, the activity is low, and the impact of the marker on the human body is still unknown

The current mainstream separation methods are not suitable for medical clinical use due to various problems such as high cost, low activity, high contamination rate, and markers.

Images