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Perfusion culturing method of hemopoietic cell in agitation type bioreactor

A bioreactor and hematopoietic cell technology, applied in biochemical equipment and methods, blood/immune system cells, microorganisms, etc., can solve the problems of differences in the degree of expansion, inability to culture a large number of cells, and low culture efficiency, and achieve Detect and implement online monitoring, facilitate online monitoring, and cultivate the effect of constant environment

Inactive Publication Date: 2004-12-22
上海伯瑞生物技术发展有限公司 +1
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AI Technical Summary

Problems solved by technology

[0006] The above inventions are all static culture systems, which have the problem of inhomogeneous culture environment. There are concentration gradients in pH, dissolved oxygen, and metabolites in the culture medium. Distribution, media composition, and degree of expansion of cultured samples can cause large variations between batches of culture
In addition, static culture systems such as well plates and square bottles cannot be monitored online, and data collection requires cumbersome operations
Finally, the static system is also limited by the surface area it can provide and cannot grow a large number of cells, which will cause great difficulties when scaling up
Therefore, the disadvantages of static culture are that the culture efficiency is not high, the reproducibility is poor, and it is difficult to scale up.

Method used

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  • Perfusion culturing method of hemopoietic cell in agitation type bioreactor
  • Perfusion culturing method of hemopoietic cell in agitation type bioreactor
  • Perfusion culturing method of hemopoietic cell in agitation type bioreactor

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Embodiment 1

[0032] After the umbilical cord blood was centrifuged with Ficoll gradient, the mononuclear cells were collected and washed twice with IMDM medium. IMDM is used as the basic medium for in vitro culture, and 20% fetal bovine serum (FBS) is added when used, as well as SCF (50ng / mL), IL-3 (2ng / mL), IL-6 (20ng / mL), FL (50ng / mL) mL) and TPO (20ng / mL), the umbilical cord blood mononuclear cells were suspended with the above-mentioned medium, and 1×10 6 cells / mL were inoculated into the reactor. The stirring speed of the reactor is 30r / min, the temperature is controlled at 37°C, the pH is controlled at 7.1, and the dissolved oxygen is 22% (air saturation).

[0033] On the 4th day of culture, start perfusion culture. The medium used for perfusion is the same as the medium at the time of inoculation. On the 7th day, the perfusion rate is D / V=1 / 5. After the cell culture reaches the 7th day, adjust the perfusion rate to D / V V=1 / 3.

[0034] The static culture of the square bottle was u...

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Abstract

A pouring culture method of hematopoietic cells in stirring type bioreactor includes suspending the hematopoietic cells in culture medium, inoculating to stirring-type bioreactor, culturing continuously pouring the culture medium in the bioreactor, flowing the cultured liquid in a settling device, intercepting the living cells, and returning then back to the bioreactor. Its advantage is high amplification efficiency.

Description

technical field [0001] The invention relates to a method for culturing hematopoietic cells, in particular to a method for perfusion culturing hematopoietic cells through a stirred bioreactor. Background technique [0002] Hematopoietic cells include various types of blood cells and lymphocytes. During the development of the hematopoietic system in the body, all hematopoietic cells are differentiated from hematopoietic stem cells. The in vitro culture technology of hematopoietic stem cells is based on simulating the occurrence and development of the hematopoietic system in vivo, adding hematopoietic cytokines and various nutrients, and controlling the culture conditions in the culture system, so as to realize the proliferation and development of hematopoietic cells, and the production can be used for medical treatment. Hematopoietic stem / progenitor cells or various types of mature hematopoietic cells. [0003] In hematopoietic cell culture, in order to replenish nutrients in...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/02C12N5/078
Inventor 谭文松迟占有蔡海波
Owner 上海伯瑞生物技术发展有限公司
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