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Human marrow, umbilical cord blood and peripheral blood stem cell isolation kit and isolation method thereof

A peripheral blood stem cell and separation method technology, which is applied to human bone marrow, umbilical cord blood, and peripheral blood stem cell separation kits and the field of separation thereof, can solve the problems of influence of vitality, time-consuming and expensive, cell loss, etc., and achieves safety guarantee and reduces Separation cost, simple operation effect

Inactive Publication Date: 2017-01-25
SHANGHAI JIAO TONG UNIV +1
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  • Claims
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Problems solved by technology

The whole process is time-consuming and expensive, with severe cell loss and significantly affected viability

Method used

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  • Human marrow, umbilical cord blood and peripheral blood stem cell isolation kit and isolation method thereof
  • Human marrow, umbilical cord blood and peripheral blood stem cell isolation kit and isolation method thereof
  • Human marrow, umbilical cord blood and peripheral blood stem cell isolation kit and isolation method thereof

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Embodiment

[0069] A bone marrow, umbilical cord blood, and peripheral blood stem cell separation kit include the following reagents: reagent A, reagent B, and reagent C; the reagent A is sodium chloride solution; the reagent B contains hydroxyethyl starch and mixed antibodies Sodium chloride solution; the reagent C is iodixanol working solution or lymphocyte separation solution. Wherein, in the reagent B, the mass concentration of the hydroxyethyl starch is 5-10%; the concentration of the mixed antibody is 4-12 micrograms / ml.

[0070] The preparation steps of the reagent B are as follows: take 100 microliters of the mixed antibody solution and add it to the hydroxyethyl starch physiological saline solution with a mass concentration of 5-10%, to obtain the reagent B.

[0071] The mixed antibody in Reagent B was prepared by the following method:

[0072] S1. Weigh 1 milligram of antibodies (anti-CD2, CD3, CD14, CD16, CD19, CD24, CD56 and CD66b, etc.);

[0073] S2, adding 3 mg of anti-gly...

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Abstract

The invention provides a human marrow, umbilical cord blood and peripheral blood stem cell isolation kit and an isolation method thereof. The isolation kit comprises a reagent A, a reagent B and a reagent C, wherein the reagent A is sodium chloride solution, the reagent B is sodium chloride solution with hydroxyethyl starch and a mixed antibody, and the reagent C is iodixanol operating fluid or lymphocyte separating medium. In the reagent B, a mass concentration of the hydroxyethyl starch is 5-10%, and a concentration of the mixed antibody is 4-12microgram / ml. The isolation method is characterized in that a negative selection collection method is adopted for selectively removing red blood cells, blood platelets, all plasma substances, mature blood cells and lymphocytes; all lin negative stem / progenitor cells can be collected; adoption of a mobilizing agent and plasma residuals are avoided; high-capacity isolation of marrow, umbilical cord blood and peripheral blood can be realized, and cell recovery rate reaches 85%. The isolation kit and the isolation method are especially suitable for treatment of marrow, umbilical cord blood and peripheral blood samples extracted clinically and in umbilical cord blood banks.

Description

technical field [0001] The present invention relates to a human bone marrow, umbilical cord blood and peripheral blood stem cell separation kit and a separation method thereof, in particular to a kit for rapidly separating large-capacity human bone marrow, umbilical cord blood and peripheral blood stem cells made by using the negative selection method principle, It is composed of diluent, separation liquid and washing liquid to meet the needs of clinical stem cell therapy. Background technique [0002] As 2007 came to the end of the year, the life science field was surging again. Two research groups in Japan and the United States successively used transgenic technology to introduce Oct4, nanog, sox2 and other genes into ordinary human fibroblasts, and found that the reprogramming of fibroblasts possessed the characteristics of human embryonic stem cells. The results of this research caused a shock all over the world. The U.S. government has made persistent efforts and inve...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0789C12N5/077
CPCC12N5/0634C12N5/0647C12N5/0662C12N2500/12C12N2500/34C12N2509/00
Inventor 崔大祥汪铮张倩尹婷
Owner SHANGHAI JIAO TONG UNIV
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