Recovery processing method for increasing reuse times of ultra-microcarrier

A technology of recycling and microcarriers, which is applied in the direction of artificial cell constructs, vertebrate cells, animal cells, etc., can solve the problems of less reuse times, waste, and no recycling method, so as to increase the number of times of use and save use cost effect

Active Publication Date: 2012-07-18
ZHAOQING DAHUANONG BIOLOGIC PHARMA +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Although microcarriers are widely used in the biological product industry, due to the immaturity of microcarrier manufacturing technology in my country, they cannot be applied to high-density cell culture in bioreactors.
The microcarriers used in China are all imported products, but imported microcarriers usually have the problem of high price, which has brought a great impact on the development of biological products in my country, especially the development of veterinary biological products
At the same time, most of these imported microcarriers are disposable carriers, and only a small number of microcarriers can be reused, but the number of reuses is relatively small. Give an effective recycling method
At present, the commonly used microcarrier recovery process is too simple, resulting in unclean treatment of residual cell debris on the surface of microcarriers, affecting the reuse of microcarriers; because there is no effective systematic recovery method, most of the microcarriers are not recovered after use. Recycling and reuse, resulting in serious waste

Method used

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  • Recovery processing method for increasing reuse times of ultra-microcarrier
  • Recovery processing method for increasing reuse times of ultra-microcarrier
  • Recovery processing method for increasing reuse times of ultra-microcarrier

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] 1. Carrier: Cytodex Ⅰ is from Pharmacia (Mafasia), its material is crosslink dextrin (cross-linked dextran), and its specific surface area is 270cm 2 / ml.

[0045] 2. Experiment: The specific operation is as follows:

[0046] (1) Carrier recovery: After the newly purchased Cytodex I microcarriers are used once, filter the solution with a 200-mesh stainless steel screen and place it in a container. 10mM EDTA in PBS solution, then dilute with medical alcohol to the volume concentration of the carrier to 20%, and stir at a medium speed. During the stirring process, filter out the solvent and replace the 0.1M PBS solution containing 10mM EDTA once, and continue to stir , after the stirring is finished, use a 300-mesh screen to filter and recover the microcarriers, and at the same time take out the cell fragments that have come off from the microcarriers from the solution; put the microcarriers collected after filtration into the container, and then add 1 times the volume o...

Embodiment 2

[0051] 1. Carrier: Mica is from Anawa Company, its material is Polyacrylate (polyacrylate), and its specific surface area is 130 cm 2 / ml.

[0052] 2. Experiment: The specific operation is as follows:

[0053] (1) Carrier recovery: filter the newly purchased Mica microcarriers through a 200-mesh stainless steel screen to remove the solution after one use, and place it in a container. The PBS solution of EDTA is diluted with medical alcohol to the volume concentration of the carrier to 20%, and stirred at a medium speed. During the stirring process, the solvent is removed by filtration and replaced with a 0.1M PBS solution containing 50mM EDTA once, and the stirring is continued. After the stirring, use a 300-mesh sieve to filter and recover the microcarriers, and at the same time take out the cell fragments from the microcarriers from the solution; put the microcarriers collected after filtration into the container, and then add 1 times the volume of the microcarriers The co...

Embodiment 3

[0057] 1. Carrier: Biosilon, from Nunc Company, its material is Polystyrene (polystyrene), and its specific surface area is 160 cm 2 / ml.

[0058] 2. Experiment: The specific operation is as follows:

[0059] (1) Carrier recovery: Filter the newly purchased Biosilon microcarriers through a 100-mesh stainless steel screen to remove the solution after one use, and place it in a container. The PBS solution of EDTA is diluted with medical alcohol to the volume concentration of the carrier to 20%, and stirred at a medium speed. During the stirring process, the solvent is removed by filtration and the PBS solution containing 0.1M EDTA with a concentration of 10mM is replaced once, and the stirring is continued. After the stirring, use a 200-mesh sieve to filter and recover the microcarriers, and at the same time take out the cell fragments from the microcarriers from the solution; put the microcarriers collected after filtration into the container, and then add 1 times the volume o...

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Abstract

The invention relates to a recovery processing method for a microcarrier, in particular to a recovery processing method for increasing reuse times of an ultra-microcarrier, which includes steps of screening, diluting a solution, stirring, filtering, processing by using sodium hydroxide, adjusting pH value, processing of enzymic digestion, washing, processing of sterilizing and the like. The recovery processing method for increasing reuse times of the ultra-microcarrier adopts combination of a plurality of steps to achieve a best processing effect, remarkably increases use times of the microcarrier, and saves use cost. The microcarrier after being processed for a plurality of times does not have remarkable effect on wall-attaching growing conditions in the microcarrier of a cell during use. The processing method can be widely applied to recovery of various microcarriers.

Description

[0001] technical field [0002] The invention relates to a recycling and processing method for microcarriers, in particular to a recycling and processing method for increasing the number of times the microcarriers are reused. [0003] Background technique [0004] Large-scale in vitro culture technology to cultivate mammalian cells is an effective method for the production of biological products. In the past decade or so, the human demand for biological products such as growth hormone, interferon, monoclonal antibody, vaccine and interleukin has soared. At present, animal cells that can be cultured on a large scale include chicken embryos, monkey kidneys, hamster kidneys and other primary cells and human diploid cells, such as BHK-21 (hamster kidney cells), MDCK, Marc-145, etc., and have been Successfully produced products including rabies vaccine, foot-and-mouth disease vaccine, swine fever and blue ear disease vaccine, etc. Attitudes toward animal cell culture have now ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/071
Inventor 徐家华陈瑞爱施维松汤钦张东霞
Owner ZHAOQING DAHUANONG BIOLOGIC PHARMA
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