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Double-loop oscillation perfusion-type biological reaction system

A biological reaction and perfusion technology, applied in the field of biomedical engineering, can solve the problems of increasing production costs, lack of universality, and increasing production costs, and achieve the effects of saving production costs, reducing the risk of pollution incidents, and reducing consumption

Inactive Publication Date: 2013-07-10
NANFANG HOSPITAL OF SOUTHERN MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, how to minimize the non-perfusion path and ineffective circulation and improve the perfusion efficiency has always been a problem that needs to be broken through; the stress stimulation of a single plane in perfusion culture is difficult to imitate the in vivo mechanical environment of cells is also a recent research hotspot, even for biomaterials Advanced pretreatment can improve the construction efficiency of engineered tissues, but invisibly increases the production cost, which is not universal; moreover, repeated perfusion of the same culture medium will inevitably lead to the accumulation of metabolic waste, and continuous perfusion of new culture medium, Discarding the culture medium containing metabolic waste will greatly increase the production cost. How to obtain high-efficiency and high-quality engineered tissues at low cost is a problem that needs to be solved in the field of biomedical engineering

Method used

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  • Double-loop oscillation perfusion-type biological reaction system
  • Double-loop oscillation perfusion-type biological reaction system
  • Double-loop oscillation perfusion-type biological reaction system

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Example 1: Construction of the dual-loop oscillating perfusion bioreactor system of the present invention.

[0042] The dual-loop oscillating perfusion bioreaction system described in the present invention is suitable for different types of seed cells (such as cells from various sources such as human, mouse, rabbit, sheep, monkey, etc.) and biological scaffold materials with certain biomechanical strength. co-cultivate.

[0043] Double-loop oscillating perfusion bioreactor system, such as figure 1 As shown, it comprises: a support perfusion culture device 14, which contains a plurality of supports 13 for co-cultivation of seed cells and biological materials; the top surface of the support perfusion culture device 14 is equipped with a liquid inlet pipe 1, and the bottom surface The liquid outlet pipe 7; the liquid inlet pipe 1 and the liquid outlet pipe 7 are respectively connected to the infusion tube; the liquid storage bottle 11 is used to transport the culture so...

Embodiment 2

[0045] Example 2: Design and assembly of scaffold perfusion culture device.

[0046] The scaffold perfusion culture device is the key device of the dual-loop oscillation perfusion bioreactor system of the present invention, and its design and assembly are described in detail as follows.

[0047] The scaffold perfusion culture device, such as Figure 2 to Figure 5 As shown, it includes: a perfusion body 4, which is provided with a plurality of perfusion holes 41 in the liquid perfusion direction, and at least one bracket can be accommodated in the perfusion holes 41; the perfusion device upper cover 6, whose top surface is a smooth plane, and its bottom surface is opened There is a groove that can partially accommodate the perfusion body; the upper cover 6 of the perfusion device is provided with a liquid inlet hole that can be inserted into the liquid inlet pipe 1; the bottom surface of the perfusion device 2 is a smooth plane, and the top surface is opened There is a groov...

Embodiment 3

[0053] Example 3: Isolation, cultivation and identification of seed cells (New Zealand white rabbit costal chondrocytes).

[0054] Two- to three-month-old New Zealand white rabbits were sacrificed by air embolism of the ear veins. The bilateral thorax was exposed under aseptic conditions, and the costal cartilages on both sides were removed. Use PBS containing 1% double-antibody (penicillin, streptomycin) in the ultra-clean bench to wash away the blood stains, cut off the outer residual muscle and the dense perichondrium surrounding the cartilage, cut the cartilage into pieces, and wash with 0.25% trypsin at 37°C Digest for 30min, centrifuge at 1000rpm for 5min, discard the supernatant, add 0.2% type II collagenase for digestion for 6-8h, centrifuge at 1000rpm for 10min, rinse the precipitate with DMEM. Inoculate the obtained costal chondrocytes on 75cm 2 In the culture flask, add DMEM containing 10% fetal bovine serum and 1% double antibody, and place at 37°C with 5% CO 2...

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Abstract

The invention relates to a double-loop oscillation perfusion-type biological reaction system which is applied to co-culture of seed cells and biomaterials. The biological reaction system comprises a support perfusion culture device, a liquid storage bottle, a recovery bottle and peristaltic pumps, wherein the support perfusion culture device is internally provided with a plurality of supports for co-culture of seed cells and biomaterials; the liquid storage bottle is used for conveying a culture solution in the bottle to the supports; the recovery bottle is used for receiving the culture solution flowing from the support perfusion culture device through a liquid conveying pipe; the flowing direction of the liquid storage bottle and the flowing direction of the recovery bottle are controlled through respective three-way pipes, so that the liquid storage bottle and the recovery bottle are switched between a perfusion liquid-changing channel and a perfusion culture channel; the peristaltic pump is respectively arranged in the perfusion liquid-changing channel and the perfusion culture channel and used for controlling the flow velocity of a culture solution passing through the liquid conveying pipe. According to the double-loop oscillation perfusion-type biological reaction system, the combination between an oscillation method and the perfusion method is achieved, the seed cells inside the biomaterials are subjected to sufficient substance interchange and stress stimulation, the production efficiency of engineered tissues is improved and the requirements of large-scale classed culture can be met.

Description

technical field [0001] The invention belongs to the field of biomedical engineering, and relates to a bioreactor for constructing artificial tissues or organs using tissue engineering technology, in particular to a double-loop oscillating perfusion bioreactor system applied to the co-cultivation of seed cells and biological materials. Background technique [0002] Current tissue engineering research mainly includes four aspects: seed cells, biomaterials, in vivo and in vitro construction of engineered tissues or organs, and clinical application of tissue engineering. The core of tissue engineering is to establish a three-dimensional space complex composed of cells and biomaterials to simulate the three-dimensional living environment of cells in the body, which is more conducive to the exertion of the essential characteristics of cells, which is different from the traditional two-dimensional culture (cell culture) method. essential difference. With the rise of three-dimensi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12M3/00
Inventor 金丹夏立恒余斌郭小磊王磊周健张永涛梅刚
Owner NANFANG HOSPITAL OF SOUTHERN MEDICAL UNIV
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