Indirect contact co-culture system capable of realizing engineering amplification

A co-culture system and engineering technology, applied in general culture methods, tissue cell/virus culture devices, tissue culture, etc., can solve problems such as difficult to achieve engineering amplification, unable to meet the production needs of cell products, etc., to achieve the benefits of cell Effects on Physiological Function

Inactive Publication Date: 2018-06-01
DALIAN UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The biggest problem with these two culture methods is that their indirect contact co-culture system can only be used for laboratory research, it is difficult to achieve engineering scale-up, and cannot meet the production needs of cell products

Method used

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  • Indirect contact co-culture system capable of realizing engineering amplification
  • Indirect contact co-culture system capable of realizing engineering amplification

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] 【1】Prepare sodium alginate solution with a concentration of 30g / L;

[0031] [2] Prepare a collagen solution with a concentration of 5g / L and adjust it to neutral with NaOH;

[0032] [3] Prepare the gel bath solution, which contains calcium chloride (concentration 7g / L), sodium chloride (concentration 3g / L), sodium dihydrogen phosphate (concentration 1g / L) and F68 solution (1g / L) in the gel bath. L);

[0033] 【4】Preparation of chitosan solution, the concentration is 5g / L;

[0034] [5] Mix the sodium alginate solution prepared in step [1] with the collagen solution prepared in step [2] in a volume ratio of 2:1;

[0035] [6] Collect the hepatic parenchymal cells cultured to the logarithmic growth phase by centrifugation, and evenly disperse them into the mixed solution prepared in step [5] at a density of 1×106 / ml;

[0036] [7] The mixture of sodium alginate and collagen containing hepatic parenchymal cells prepared in step [6] forms a stable jet under a high-voltage el...

Embodiment 2

[0049] 【1】Prepare sodium alginate solution with a concentration of 40g / L;

[0050] [2] Prepare a collagen solution with a concentration of 4g / L and adjust it to neutral with NaOH;

[0051] [3] Prepare the gel bath solution, which contains calcium chloride (concentration 7g / L), sodium chloride (concentration 3g / L), sodium dihydrogen phosphate (concentration 1g / L) in the gel bath;

[0052] 【4】Preparation of chitosan solution, the concentration is 5g / L;

[0053] [5] Mix the sodium alginate solution prepared in step [1] with the collagen solution prepared in step [2] in a volume ratio of 1:2;

[0054] 【6】The fetal mouse fibroblasts cultured to the logarithmic growth phase were collected by centrifugation, according to 2×10 5 The density of / ml is uniformly dispersed in the mixed solution prepared in step [5];

[0055] [7] The mixture of sodium alginate and collagen containing fetal mouse fibroblasts prepared in step [6] and the gel bath solution prepared in step [3] were passed...

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Abstract

The invention discloses an indirect contact co-culture system capable of realizing engineering and large-scale amplification as well as a preparation method thereof. The system consists of a bioreactor capable of containing numerous tiny unit particles. The tiny unit particles are spherical hydrogel carriers with the particle size of 100 to 1000 microns, and the surface of each hydrogel ball has one layer of hydrogel membrane structure. One kind of cells are in the spherical hydrogel, the other kind of cells are distributed on the surface of the spherical hydrogel membrane, the two kinds of cells are formed and can cause cell membrane contact through the membrane pores of the hydrogel membrane, and the cells which are separated at downstream can be in direct contact with a co-culture unitconveniently.

Description

technical field [0001] The invention relates to an indirect contact co-cultivation system, in particular to an indirect contact co-cultivation system characterized by a spherical membrane. Background technique [0002] Cell co-culture technology provides the possibility for the in vitro study of cell biology. According to the degree of contact between co-cultured cells, the current co-culture technology is divided into three types: direct contact co-culture, non-contact co-culture and indirect contact co-culture. Among them, the direct contact co-culture technology is beneficial to the function of the cells due to the full contact between the co-cultured cells, but the disadvantage is that after the co-cultivation is over, it is difficult to separate the two cells, and only one of the cells can be fluorescently labeled. Separation by flow sorting, or by magnetic field separation after one of the cells has been labeled with immunomagnetic beads, is tedious and expensive. In...

Claims

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Application Information

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IPC IPC(8): C12M3/00C12N5/00
CPCC12M25/16C12M25/18C12M25/20C12N5/0075C12N2533/52C12N2533/54C12N2533/70C12N2533/72C12N2533/74
Inventor 于炜婷刘袖洞梁珊珊王若雨马小军
Owner DALIAN UNIVERSITY
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