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Method and device for perfusion and culture of hematopoietic cell

A technology for perfusion culture and hematopoietic cells, applied in tissue cell/virus culture devices, tissue culture, biochemical cleaning devices, etc., can solve problems such as concentration gradients, uneven culture environments, cumbersome data collection operations, etc., to achieve high Amplification multiple, avoiding the effect of cells not growing or growing slowly

Inactive Publication Date: 2004-12-29
上海伯瑞生物技术发展有限公司 +1
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  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the static culture system has problems such as inhomogeneous culture environment, concentration gradients, etc., and there are shortcomings such as difficulties in online monitoring and cumbersome data collection operations.

Method used

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  • Method and device for perfusion and culture of hematopoietic cell
  • Method and device for perfusion and culture of hematopoietic cell
  • Method and device for perfusion and culture of hematopoietic cell

Examples

Experimental program
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Effect test

Embodiment 1

[0061] Structural parameters of the reactor: height = 400mm, diameter = 100mm; the stirring paddle 27 adopts a 30° oblique blade lifting type stirring paddle, and the ratio of the diameter of the paddle 27 to the diameter of the container 2 is 4:5.

[0062] The perfusion culture chamber 102 is a square tube with a water bath jacket outside, and its dimensions are 1.2 cm high, 1.5 cm wide, and 20 cm long;

[0063] Training method:

[0064] After the umbilical cord blood was centrifuged with Ficoll gradient, the mononuclear cells were collected and washed twice with IMDM medium. The basic medium used for in vitro culture was IMDM, and 20% fetal bovine serum (FBS), 50 ng / mL of SCF, 10 ng / mL of IL-3, 20 ng / mL of IL-6, 50 ng / mL of FL and 25ng / mL TPO, suspend cord blood mononuclear cells with the above medium, 1×10 6 cells / mL were inoculated into the culture chamber, and the temperature of the water bath was controlled at 37°C. The temperature, pH, and dissolved oxygen of the cul...

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Abstract

The present invention is perfusion culture medicine method and apparatus for hematopoietic cell. The apparatus includes at least stirring bioreactor and perfusing culture cavity connected via pipeline to the stirring bioreactor. The perfusion culture process includes first static perfusion culture until reaching higher cell density, and subsequent stirring suspension culture to eliminate the cell density gradient and accelerate cell growth. Test result shows that the initial culture period has cloning in great amount of total cell as well as CFU-Mix, CFU-GM and CD34+ cell, and the subsequent stirring suspension culture stage has further cloning of total cell as well as CFU-Mix, CFU-GM and CD34+ cell. The said method has greater cloning than single stirring suspension culture.

Description

technical field [0001] The invention relates to a hematopoietic cell perfusion culture method and device. Background technique [0002] In the in vitro culture system of hematopoietic cells, perfusion culture can replenish cytokines and nutrients in time and remove metabolic byproducts. Therefore, compared with pure static fed culture, it can not only increase the expansion of hematopoietic cells, but also make LTC-IC was amplified. The current perfusion culture system is generally a static perfusion chamber reactor, such as Sandstrom CE, Bender JG, Papoutsakis ET, Miller WM. Effects of CD34 + Cell selection and perfusion on ex vivo expansion of peripheral blood mononuclear cells.Blood.1995,86(3):958-70. The reported perfusion culture system without stromal cells, and Koller MR, Manchel I, Maher RJ, et al .Clinical-scale human umbilical cord blood cell expansion in a novel automated perfusion culture system. Bone Marrow Transplant.1998, 21 (7): 653-63. The reported automa...

Claims

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Application Information

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IPC IPC(8): C12M1/02C12M3/02C12N5/08
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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