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Method and preparation for preparing mature erythrocytes from peripheral blood in vitro

A technology for mature red blood cells and peripheral blood, applied in cell dissociation methods, cell culture active agents, biochemical equipment and methods, etc.

Active Publication Date: 2020-01-03
WEST LAKE BIOMEDICAL TECH HANGZHOU CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The shortage of clinical blood has always been a problem in the national and global medical and health fields. According to research, the amount of clinical blood used in medical care is increasing at a double-digit rate year by year. At the same time, the number of qualified blood donors is decreasing. Coupled with the cancellation of mutual aid blood donation in some parts of our country, it has led to a serious contradiction between blood supply and demand-supply exceeds demand; and it cannot be ignored that there is still a risk of blood-borne disease transmission through allogeneic blood transfusion, due to the "window period" of blood "Many new pathogens will not be detected during the detection period, which brings a certain degree of risk of blood transfusion
In addition, blood transfusion medicine is also faced with the problems of antigen / antibody mismatch leading to immune resistance in patients and difficulty in obtaining suitable blood sources in time for patients with rare blood types. Nervous questions became looming conundrums
[0003] In order to solve the above blood problem, red blood cell regeneration technology has developed rapidly. The current red blood cell regeneration method is mainly based on CD34 + Bone marrow hematopoietic stem cells or CD34 + Umbilical cord blood hematopoietic stem cells, although this scheme can also achieve erythroid hematopoietic differentiation in vitro, but the extraction of CD34 + Bone marrow hematopoietic stem cells can cause great physical pain and harm to cell suppliers, while CD34 + Umbilical cord blood hematopoietic stem cells can only be obtained during a specific period of time after the birth of the newborn, and it is difficult to obtain the cells. Therefore, the above two methods have great difficulties in clinical application.
[0004] The Chinese patent "A Method for Preparing Mature Red Blood Cells" with the authorization number CN 103667188B mentions the method of using mononuclear cells in umbilical cord blood or peripheral blood to prepare mature red blood cells through in vitro culture. A method of directly preparing red blood cells from blood using mononuclear cells as starting materials, but there are problems in this method: it is still through the amplification of CD34 + It is very difficult to obtain red blood cells by means of cells, and the possibility of large-scale clinical application is small

Method used

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  • Method and preparation for preparing mature erythrocytes from peripheral blood in vitro
  • Method and preparation for preparing mature erythrocytes from peripheral blood in vitro
  • Method and preparation for preparing mature erythrocytes from peripheral blood in vitro

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0069] 1. Ingredients preparation:

[0070] 1.1 Biotin-labeled antibody

[0071] To prepare 1ml of biotin-labeled antibody, the following components are required:

[0072] Element volume Biotin-labeled mouse anti-human CD3 antibody 20ul Biotin-labeled mouse anti-human CD14 antibody 20ul Biotin-labeled mouse anti-human CD16 antibody 20ul Biotin-labeled mouse anti-human CD19 antibody 20ul Biotin-labeled mouse anti-human CD41a antibody 20ul Biotin-labeled mouse anti-human CD56 antibody 20ul Biotin-labeled mouse anti-human CD235a antibody 20ul

[0073] 1.2. Enriched cell culture medium (including hematopoietic stem cell expansion medium, cytokine combination and penicillin and streptomycin)

[0074] To configure 1L enriched cell culture medium, the following components are required:

[0075]

[0076]

[0077] 1.3. Configuration of medium for the first stage of differentiation:

[0078] To configure 1L of the first-...

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Abstract

The invention relates to a method and preparation for preparing mature erythrocytes in vitro by utilizing peripheral blood, in particular to a method for utilizing Lin in peripheral blood ‑ CD34 ‑ Method for preparing mature erythrocytes from cells, including enriching Lin ‑ CD34 ‑ Cells that differentiate into erythroid cells Lin ‑ CD34 ‑ cells and the step of promoting the denucleation and maturation of red blood cells; in addition, it also involves the mature red blood cells prepared by the method and the differentiation medium and cytokine combination used therein. The preparation method has the following advantages: the raw materials are easy to obtain, the yield of red blood cells is high, The resulting erythrocytes performed well.

Description

technical field [0001] The invention relates to the field of stem cells and cell regeneration, in particular to a method and method for preparing mature red blood cells in vitro by using peripheral blood, the mature red blood cells prepared by the method, and the use of mononuclear cells for preparing mature red blood cells. Background technique [0002] It is well known that blood transfusion is a life-saving means to treat patients with severe ischemia (such as acute anemia, severe thrombocytopenia, hemophilia), but there are many problems in clinical medicine, mainly reflected in the two aspects of blood shortage and blood transfusion risk. aspect. The shortage of clinical blood has always been a problem in the national and global medical and health fields. According to research, the amount of clinical blood used in medical care is increasing at a double-digit rate year by year. At the same time, the number of qualified blood donors is decreasing. Coupled with the cancel...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/078
CPCC12N5/0641C12N2501/125C12N2501/14C12N2501/2306C12N2501/2303C12N2509/00C12N2501/998C12N2501/727C12N2501/515C12N2501/50C12N2501/599C12N2500/32C12N2500/24C12N2501/33C12N2500/90C12N2500/25C12N2501/26C12N2506/115
Inventor 高晓飞黄彦杰张清
Owner WEST LAKE BIOMEDICAL TECH HANGZHOU CO LTD
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