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Rotary cell-culture system

A cell culture, rotary technology, applied in tissue cell/virus culture devices, biochemical instruments, biochemical equipment and methods, etc., to improve the culture effect, prevent the entry of air bubbles, and avoid damage.

Inactive Publication Date: 2004-11-03
DALIAN UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

It reduces the shear force of the cell growth environment, improves the material transfer efficiency, and effectively overcomes the large shear force on the cells in the existing bioreactor, the instability of the cell growth environment, and the fact that the reactor must be placed in CO 2 Insufficient technology such as operation in the incubator

Method used

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Examples

Experimental program
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Effect test

Embodiment 1

[0029] This example is the result of microcarrier culture of osteoblasts in this system.

[0030] Preparation of seed cells: under aseptic conditions, take the calvaria of SD rats born 3 days old, put them into a culture dish filled with buffer solution, remove the periosteum and connective tissue, and rinse repeatedly. The washed calvaria slices were placed in a petri dish, added with 0.25% trypsin solution, and digested at a constant temperature of 37°C for 15-20min. Then the calvarial slices were digested with 1mg / ml type II collagenase solution at 37°C for 90min. Dissociate cells from bone matrix. Transfer the digestion solution to a centrifuge tube and centrifuge for 10 min (1000 rpm) to pellet the cells. The supernatant was discarded, and the precipitate was washed twice with RPMI1640 culture medium. Then pipette the bone block to obtain more osteoblasts. Repeat the centrifugation process to pellet the cells and make a cell suspension. Cells were inoculated into cul...

Embodiment 2

[0035] This example is the result of culturing hematopoietic stem cells in this system.

[0036] Preparation of umbilical cord blood hematopoietic stem cells: cord blood samples are taken from placental tissue of normal delivered full-term infants. About 60-80 ml of blood is drawn from the umbilical vein with a blood collection needle and injected into a heparin anticoagulated centrifuge tube. Then, the cord blood was evenly diluted with an equal volume of IMDM medium, centrifuged and sedimented to discard the fat-containing turbid liquid, and then diluted with IMDM. Then suck the cell suspension, carefully add it to the surface of an equal volume of lymphocyte separation solution along the tube wall, centrifuge at 1500rpm for 20min, collect the leukocyte layer, add IMDM solution for repeated washing, and then centrifuge at 1000rpm for 10min. Umbilical cord blood CD was isolated and purified by immunomagnetic strain method. 34 + Hematopoietic stem cells.

[0037] Cultured i...

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Abstract

The present invention relates to biological technology, and is one kind of rotary cell culture system as large-scale animal cell culturing system. Technologically, the rotary cell culture system includes rotary cell culture reactor, culture medium oxygen inflator, outer culture medium circulating system and electric control system. It has penetrating oxygen supplier and two, one outer and one inner, rotating reactor barrels, which can be regulated in speed alone for changing liquid continuously or intermittently and may be operated outside the CO2 culture case. It is superior to available bioreactor with great cell shearing force, instable cell growth environment and operation inside the CO2 culture case. The present invention can make osteoplast and hemopoietic stem cell well grow to result in cell proliferation of 23 times and 49 times separately in 10 days, and is suitable for large scale culture of cells.

Description

technical field [0001] The invention belongs to the field of biotechnology, in particular to a rotary cell culture system for large-scale culture of animal cells. Background technique [0002] So far, animal cell culture can be divided into static two-dimensional culture and dynamic three-dimensional culture. The former is mainly suitable for small-scale research in the laboratory, while large-scale cultivation of animal cells must rely on biotechnology under three-dimensional conditions. Traditional cell culture systems mainly include air-lift type, stirring type, etc. The main problem of these types of culture systems is that the cells are subjected to a large fluid shear force in the reactor, making cells that are sensitive to shear force very easy to Damaged, especially all kinds of stem cells. Therefore, cells that are extremely sensitive to shear stress, such as stem cells, are difficult to be cultured and expanded on a large scale in these traditional cell culture s...

Claims

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Application Information

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IPC IPC(8): C12M3/00
Inventor 孙相彧刘天庆李香琴
Owner DALIAN UNIV OF TECH
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