Device and method for in vitro high-density cultivation of erythrocyte

Abstract

The invention discloses a device for in vitro high-density cultivation of erythrocyte. The device comprises a stirred tank bioreactor and a hollow fiber cell intercepting device, wherein the structure of the hollow fiber cell intercepting device includes a cylindrical sleeve formed by 4-10 hollow fiber tubes which are parallel to one another; an elastic valve, a piston and a vacuum pump are orderly arranged at the bottom of the cylindrical sleeve; and the upper part of the cylindrical sleeve is connected with the stirred tank bioreactor through a connecting pipe. A method for in vitro high-density cultivation of erythrocyte is also disclosed by the invention. The method comprises the following steps of (1) separating a mononuclear cell; (1) separating CD34+ hematopoietic stem cell; and (3) differentiating the hematopoietic stem cell into the erythrocyte. By using the device and the method, hematopoietic stem cells with different sources can be utilized to carry out in vitro differentiation, so as to form a lot of erythrocytes with complete functions. High-density cultivation of the erythrocytes can be achieved; and the density is improved by about 200 times in comparison with a common cell cultivation method.

Description

technical field [0001] The invention relates to a device and method for in vitro high-density culture of red blood cells, belonging to the field of regenerative medicine. Background technique [0002] At present, the five major problems that need to be solved in the blood transfusion system relying on artificial blood donation are: 1) The demand for blood is constantly increasing; 2) The aging society has led to a continuous decline in the number of blood donors; 3) Blood bias, such as rare blood types, it is difficult to find a match; 4) Difficult blood matching, such as anti-erythrocyte multiple immunity and other phenomena; 5) Blood transfusion safety issues. [0003] The production of mature red blood cells from hematopoietic stem cells for blood transfusion seems to provide a possible solution to this worldwide problem. Blood, fetal liver, umbilical cord blood) successfully induced mature red blood cells in vitro, and experiments in mice showed that these red blood cel...

AI Technical Summary

Problems solved by technology

[0002] At present, the five major problems that need to be solved in the blood transfusion system relying on artificial blood donation are: 1) The demand for blood is constantly increasing; 2) The aging society has led to a continuous decline in the number of blood donors; 3) Blood bias, such as rare blood types, it is difficult to find a match; 4) Difficult blood matching, such as anti-erythrocyte multiple immunity and other phenomena; 5) Blood transfusion safety issues

[0004] Although the induction of erythrocytes in vitro provides a powerful weapon to solve the problem of clinical blood transfusion, there are still many technical difficulties to be overcome in the real realization of industrialization and large-scale in vitro hematopoiesis. One of the important problems is the high-density culture technology in vitro, because a Unit red blood cell content is about 2x10 12 Red blood cells, if the conventional cell culture density (1 ~ 2 × 10 6 cell / ml), the volume needed to cultivate each unit of red blood cells is very large (if the culture bottle is tiled, the area is about 4 tennis courts), it is difficult to meet the needs of industrial production, and there is an urgent need for a method that can increase the cell culture density. Apparatus and method for reducing production space to meet the needs of large-scale production

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