Perfusion Bioreactor System

A bioreactor and perfusion technology, which is applied in the field of biomedical engineering, can solve the problems of inability to meet the complex and cultivation of large biomaterials and seed cells, inapplicable cell and tissue culture, and affect the normal growth of cells, etc., to achieve uniform cell growth. Planting effect, uniform distribution of mechanical stimuli, effect of maintaining activity and function

Inactive Publication Date: 2011-12-28
FOURTH MILITARY MEDICAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Its disadvantages are: due to the large shear stress generated by the rotation of the stirrer, it is easy to generate a large number of air bubbles, so it is not suitable for the cultivation of cells and tissues in tissue engineering
Its disadvantages are: small operating flexibility, poor mixing performance at low flow rates, especially when the reactor is high, large in diameter, and high-density microcarrier culture
The disadvantage of rotating wall bioreactor is that it is difficult to scale up industrially, and there are always s

Method used

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Experimental program
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Effect test

Embodiment 1

[0041] Embodiment 1: Construction of the perfusion bioreaction system of the present invention

[0042] The perfusion bioreactor of the present invention mainly includes a circulation perfusion system, a constant temperature system, a gas exchange system and an on-line monitoring system, and is suitable for different types of seed cells (for example, various species of human, mouse, rabbit, sheep, monkey, etc.) The co-cultivation of stem cells) and bio-scaffold materials with certain biomechanical strength.

[0043] The whole reactor system forms a whole under the connection of medical silicone tube 90, powered by a two-way peristaltic pump 20, and the fresh culture solution is continuously pumped out from the liquid storage bottle 30, passed through the constant temperature water bath device 40, heated to 37°C, and flowed Through the monitoring probe 55, the pH value, temperature, dissolved oxygen and fluid velocity of the culture solution in the reactor are monitored in real...

Embodiment 2

[0068] Embodiment 2: Design and assembly of the training cabin

[0069] Cultivation cabin is the key device of the perfusion bioreactor of the present invention, is described in detail as follows:

[0070] Such as Figure 4As shown, the culture device includes: a base 2, which is equipped with a liquid inlet pipe interface 1 outward; a top cover 9, which is equipped with a liquid outlet pipeline interface 12; a front cover 8, which is placed on the top cover 9; 4. Set between the base 2 and the front cover 8; the bracket sleeve 5 is set in the shell of the cultivation cabin 4, one end of the bracket sleeve 5 is connected to the base 2 through the base sealing ring 3, and the other end is through the top cover The sealing ring 10 is connected to the top cover 9; the support sleeve 5 is provided with a support sleeve 7 and a sleeve sealing ring 6, so that a space 15 for accommodating biological materials is defined in the support sleeve 5; and a fixing rod 13, The two ends are...

Embodiment 3

[0081] Example 3: Isolation, cultivation and identification of seed cells (SD rat bone marrow mesenchymal stem cells).

[0082] 2-week-old SD rats were killed by neck dislocation, soaked in 75% alcohol for 15 minutes, the femur and tibia of the rat were aseptically taken out in an ultra-clean workbench, the soft tissue attached to the bone was removed, and the medullary cavity was washed 1-2 times with sterile normal saline , collected directly in a centrifuge tube. Centrifuge at 1000r / min for 10min, discard the supernatant, resuspend and count using SD rat BMSCs standard culture medium (Saiye Company, USA), and plant in 25cm 2 culture flask at 37°C, 5% CO 2 cultured in a constant temperature incubator. Change the medium after 24 hours, and then change the medium every 2-3 days. When reaching 80%-90% confluence (primary culture for 3-5 days), use 0.25% trypsin to digest and subculture, and culture in groups. The well-grown third generation cells were selected for morpholog...

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Abstract

The invention relates to a perfusion bioreactor system which comprises a circulating perfusion system, a constant temperature system, a gas exchange system and an on-line monitoring system, wherein the circulating perfusion system comprises a culture chamber, a liquid storage bottle and a peristaltic pump which are sequentially connected through a pipeline to form a perfusion culture loop; the constant temperature system comprises a constant temperature water bath device of which the inside is provided with a circuitous circulating pipeline, and is applicable to an electric heating and refrigerating control part; the gas exchange system is composed of a gas filter, a gas flowmeter, a high-pressure gas bottle, a pressure relief valve and a connecting pipeline; and the on-line monitoring system is composed of a monitoring probe, a monitor terminal, monitoring software, a data conversion chip and a mass transfer regulator. The perfusion bioreactor system provided by the invention can maximally simulate the in-vivo microenvironment in the tissue or organ in-vitro construction process, ensures sufficient substance exchange and normal physiological functions of seed cells in the biological material, enhance the yield of the seed cell/biological stent material, shorten the culture period and lower the culture cost.

Description

technical field [0001] The invention belongs to the field of biomedical engineering, and relates to a bioreactor for constructing an artificial organ with a tissue engineering method, in particular to a perfusion bioreactor system applied to inoculation and co-cultivation of seed cells and biological materials. Background technique [0002] Current tissue engineering research mainly includes four aspects: seed cells, biomaterials, methods and techniques for constructing tissues and organs, and clinical applications of tissue engineering. The core of tissue engineering is to establish a three-dimensional space complex composed of cells and biological materials, which is essentially different from the traditional two-dimensional structure (cell culture). With the rise of three-dimensional cell culture technology, more and more research focuses on the inoculation and co-cultivation of seed cells and biological materials, and a series of breakthroughs have been made. The concept...

Claims

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Application Information

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IPC IPC(8): C12M3/00
Inventor 裴国献王磊金丹江汕姜晓锐张鑫鑫李方国
Owner FOURTH MILITARY MEDICAL UNIVERSITY
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