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

Abstract

The invention relates to a method and preparation for preparing mature erythrocytes in vitro through peripheral blood, in particular to a method for preparing the mature erythrocytes through Lin-CD34-cells in the peripheral blood. The method includes the steps of enriching the Lin-CD34-cells, differentiating the Lin-CD34-cells into erythroid cells and promoting denucleation and maturation of erythrocytes. In addition, the invention also relates to the mature erythrocytes prepared by the method, a differentiated culture medium and a cytokine combination, wherein the differentiated culture medium and the cytokine combination are applied in the mature erythrocytes. The method for preparation has the advantages that raw materials are easily obtained, the yield of the erythrocytes is high, andthe performance of the obtained erythrocytes is great.

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...

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

Images