In vitro reconstruction human red blood cell substitute and preparation method thereof

A technology of human erythrocytes and substitutes, which is applied in the field of erythrocytes substitutes, and can solve problems such as hemolysis reaction and toxic phagocyte phagocytosis

Active Publication Date: 2019-04-26
CHONGQING UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At the same time, routine human allogeneic blood transfusion faces the following insurmountable shortcomings: (1) Time-consuming cross-matching is required before blood transfusion, and there is a risk of hemolytic reaction caused by blood type inconsistency; (2) Blood transfusion-related diseases, such as AIDS (HI

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  • In vitro reconstruction human red blood cell substitute and preparation method thereof
  • In vitro reconstruction human red blood cell substitute and preparation method thereof
  • In vitro reconstruction human red blood cell substitute and preparation method thereof

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Embodiment Construction

[0028] The present invention is further illustrated by the following examples.

[0029] 1. Vector preparation

[0030] Wash the whole blood with isotonic phosphate buffer, separate the blood, remove plasma, white blood cells, platelets, etc., and obtain red blood cells; then place the red blood cells in the permeabilization solution for treatment; the volume ratio of red blood cells and permeation solution is 1:1 ~1:50; the processing condition is 4°C, 1min~120min; after that, high-speed refrigerated centrifugation is carried out, the centrifugation condition is: 4~60min, 4°C, 5000rpm~20000rpm, the supernatant is collected, and the blood is dissolved, and the supernatant is washed by centrifugation Colorless and transparent, the precipitate is the carrier of ghost cells.

[0031] Wherein, the preparation of membrane breaking solution is as follows: preparing membrane breaking buffer [MgCl 2 , EDTA, Tris, PMSF] (the final concentration of each substance is 1mM, 1mM, 10mM, 1mM...

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Abstract

The invention discloses an in vitro reconstruction human red blood cell substitute and a preparation method thereof. A low-seepage method is used for performing in vitro reconstruction; in a low-seepage environment (containing rich hemoglobin), temporary holes occur in cytomembrane carriers; through the inside and outside hemoglobin concentration differences in the cytomembrane carriers, the hemoglobin enters the carriers through the temporary holes; then, the temporary holes in the cytomembrane can be sealed again through the flowability of the cytomembrane; the hemoglobin entering the cellscan be coated; then, the relative loading rate of the hemoglobin of the reconstruction human red blood cells is detected by an ELIASA; the form and the size of the reconstruction human red blood cellsare observed by a phase contrast microscope; the oxygen carrying function of the reconstruction human red blood cells is detected by a blood oxygen analysis instrument. A series of reaction after theblood infusion due to immunogenicity can be reduced; the in vitro reconstruction human red blood cell substitute can be applied to clinic first aid and the like. A proper amount of 2,3-DPG, ATP and the like are added into the hemoglobin liquid for loading for regulating the oxygen carrying function of the reconstruction human red blood cells, so that the blood volume of a hypoxic-ischemic patientis improved; meanwhile, the oxygen lack state in the body of the patient is relieved.

Description

technical field [0001] The invention relates to the field of red blood cell substitutes, in particular to an in vitro reconstructed human red blood cell substitute and a preparation method thereof. Background technique [0002] In recent years, the amount of clinical blood used in clinical practice has increased rapidly all over the world, and frequent blood shortages have caused the blood shortage to reach a crisis level. In addition, under a series of special circumstances such as sudden major natural disasters, wars and terrorist incidents, when a large number of wounded need blood transfusion treatment at the same time, the contradiction between supply and demand will increase sharply. It is estimated that even in peacetime, the whole world needs about 15 million liters of fresh blood every year, and there is a clear increasing trend. The amount of blood used for medical treatment in my country is nearly 1.3 million liters every year, and it is also increasing at a rate...

Claims

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Application Information

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IPC IPC(8): A61K38/42A61K47/46A61K49/00A61P7/08A61P39/00A61P35/00
CPCA61K38/42A61K47/46A61K49/00A61P7/08A61P35/00A61P39/00Y02A50/30
Inventor 王翔赵押金候鳗陈东任扬党琦李春龚
Owner CHONGQING UNIV
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