Universal cell processing kit and application method thereof

Abstract

The invention provides a universal cell processing kit and an application method thereof, and relates to a kit for separating stem cells from human cord blood, bone marrow and peripheral blood and an application method of the kit. The kit contains a reagent A (cell diluent), a reagent B (high-molecular-weight cell precipitant) and a reagent C (medium-density layering agent). The application method of the kit comprises the steps of: firstly, uniformly mixing the cord blood, bone marrow or peripheral blood with the cell diluent, adding the high-molecular-weight cell precipitant, standing for 30 minutes, collecting the supernatant, centrifuging, collecting the concentrated solution, adding the medium-density layering agent to the concentrated solution, centrifuging for 20 minutes, collecting a karyocyte layer and washing with saline for injection 3 times. In the invention, a cell precipitant with a molecular weight of 450000-700000 is used, the survival rate of the separated karyocytes is not lower than 98%, the collection rate of karyocytes is not lower than 85%, and the method provided by the invention is simple, safe and practical, has low cost, can realize industrialized production, and is easy to popularize.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and specifically relates to a kit for separating stem cells in human umbilical cord blood, bone marrow and peripheral blood and an application method of the kit. Background technique [0002] Stem cells are cell populations existing in the normal human body, have a high degree of self-replication and differentiation capabilities, and can effectively treat various cell damage diseases. At present, there are many methods for isolating and extracting stem cells from human umbilical cord blood, bone marrow and peripheral blood, the main methods are: in vitro expansion culture, peripheral blood cell sorting apparatus, lymphocyte separation medium, etc. In vitro expansion culture has the problems of difficult control of culture conditions, unstable products, easy pollution in vitro culture, and cancer after multiple replications; the peripheral blood cell sorter is only suitable for peripheral bloo...

AI Technical Summary

Problems solved by technology

In vitro expansion culture has the problems of difficult control of culture conditions, unstable products, easy pollution in vitro culture, and cancer after multiple replications; the peripheral blood cell sorter is only suitable for peripheral blood, and only a very small amount of hematopoietic stem cells in peripheral blood For thin sternum, a mobilizing agent is required before collection, and the operation is complicated. The amount of liquid obtained in the final separation is large, and the removal of red blood cells is not complete; the lymphocyte separation liquid is mainly used to separate mononuclear cells in peripheral blood, and the lymphocytes in the separated cells are about Accounting for 90%-95%, there are only a small number of primitive cells, while lymphocytes are mature cells and do not have the characteristics of stem cells

Other methods include magnetic bead method, fluorescent labeling method, flow cytometry method, erythrocyte precipitation method with Lin antibody, and low molecular weight cell precipitation method. Some of these methods are expensive and unaffordable for patients; The conditions are high and it is difficult to produce on a large scale; some cannot effectively retain all types of stem cells; some cells have low activity, large volume of cell fluid, long separation time, and easy contamination, etc., which limit clinical application