Umbilical cord blood component synchronous separation kit and use method thereof

A synchronous separation and kit technology, applied in the field of umbilical cord blood separation, can solve the problems of underutilization, lack of separation, waste of umbilical cord blood resources, etc., and achieve the effect of promoting full utilization

Pending Publication Date: 2021-11-30
王盛
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] At present, after the cord blood is collected by the seven major umbilical cord blood hematopoietic stem cell banks in the country, the cord blood hematopoietic stem cells are extracted and preserved, and other components in the cord blood are not fully utilized. This is a great waste of cord blood resources. The main reason is that While isolating umbilical cord blood hematopoietic stem cells, there is no mature technology to simultaneously separate umbilical cord blood platelets, red blood cells, plasma and its exosomes, neutrophils and other components

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Embodiment 1: a kind of synchronous separation kit of each component of umbilical cord blood

[0036] Including No. A dilution solution, No. B1 separation solution, No. B2 separation solution, No. C washing solution, and No. D lysate; also includes consumables, including 10 50ml centrifuge tubes, 20 15ml centrifuge tubes, and 10 10ml pipettes , 5 3ml Pasteur pipettes, 175cm 2 3 cell culture flasks;

[0037] The No. A dilution is 0.9% physiological saline, in which human serum albumin is added, and the mass concentration of human albumin added to the No. A dilution is 0.75%;

[0038] The No. B1 separating liquid is prepared by the following method: 40% polysucrose liquid is prepared into a polysucrose liquid with a mass concentration of 9% with Hank's solution; then 60% compound diatrizoate meglumine The injection was formulated with Hank's solution to form a compound diatrizoate solution with a mass concentration of 34%; finally, 9% polysucrose solution and 34% compou...

Embodiment 2

[0042] Embodiment 2: a kind of synchronous separation kit of each component of umbilical cord blood

[0043] Basically the same as Example 1, the difference is that the No. A dilution is 0.9% physiological saline; the No. C washing solution is about pH 7.4 without Ca 2+ and Mg 2+ of PBS solution.

Embodiment 3

[0044] Embodiment 3: A method for synchronously separating components in cord blood with the cord blood components synchronous separation kit described in Example 1

[0045] ① Divide 100ml of voluntary donated cord blood into two 50ml centrifuge tubes a, centrifuge at 2500rpm for 10 minutes, then absorb the upper plasma layer into centrifuge tube b;

[0046] ② Add an equal volume of No. A solution to the separated centrifuge tube a and mix well, then carefully add 3ml No. B1 separation solution, 3ml B2 No. separation solution and the mixed solution in centrifuge tube a to the 15ml centrifuge tube along the wall. In the method, centrifuge at 2000rpm for 25 minutes and separate into five layers: use a Pasteur pipette to draw the first layer into the centrifuge tube c, and draw the second layer (circular milky white cell layer) mononuclear cell layer into the centrifuge tube d. Aspirate the neutrophil layer of the third layer (annular milky white cell layer) into the centrifuge t...

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Abstract

The invention discloses a kit for synchronously separating various components of umbilical cord blood. The kit comprises a diluent A, a separating medium B1, a separating medium B2, a washing solution C and a lysis solution D, the diluent A is 0.9% normal saline, the separating medium B1 and the separating medium B2 are mixed solutions formed by mixing a 9% polysucrose solution and a 34% compound meglumine diatrizoate solution according to the volume ratio of 50: 19.11 and 50: 23.28 respectively, the washing solution C is 0.9% normal saline or a PBS solution without Ca < 2 + > and Mg < 2 + >, the pH of the PBS solution is 7.4 +/-0.1, and the lysis solution D is a red blood cell lysis solution. The invention provides a technology and a kit for synchronously separating umbilical cord blood platelets, red blood cells, plasma, exosomes, neutrophils and other components, industrialization can be realized, the technical problem that other components except hematopoietic stem cells in the umbilical cord blood are not synchronously separated can be solved, and full utilization of all the components in the umbilical cord blood can be greatly promoted.

Description

technical field [0001] The invention relates to a kit and its use method, in particular to a kit for synchronous separation of components of umbilical cord blood and its use method, which is a kit capable of synchronously separating platelets, red blood cells, plasma and A test kit for granulocytes, monocytes, endothelial progenitor cells, stem cells and lymphocytes belongs to the technical field of umbilical cord blood separation. Background technique [0002] Since the world's first successful cord blood transplant in 1988, the application and research of cord blood has made great progress. The application of cord blood in the world has exceeded 80,000 cases, and the application of cord blood in my country has reached 16,000 cases. Internationally, umbilical cord blood has been used to treat more than 100 diseases, including blood diseases and non-blood diseases. But in contrast, the application of umbilical cord blood in my country is mainly for blood diseases, and the s...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0781C12N5/078C12N5/071C12N5/0783C12N5/0786C12N5/0787
CPCC12N5/0644C12N5/0641C12N5/0642C12N5/0645C12N5/0635C12N5/0636C12N5/0692C12N2509/00C12N2509/10C12N2500/90
Inventor 王盛齐湘杰
Owner 王盛
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