Protein-free culture medium for culturing microencapsulated recombinant Chinese hamster ovary (CHO) cells and preparation method thereof

A protein-free culture medium and ovarian cell technology, which is applied in the field of protein-free culture medium for microencapsulated recombinant Chinese hamster ovary cell culture, can solve problems such as high price, and achieve the effects of simple operation, low price and avoiding potential pollution.

Inactive Publication Date: 2014-12-17
DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For example, the protein-free medium CD OptiCHO produced by Gibco is expensive, 1,000 yuan per l

Method used

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  • Protein-free culture medium for culturing microencapsulated recombinant Chinese hamster ovary (CHO) cells and preparation method thereof
  • Protein-free culture medium for culturing microencapsulated recombinant Chinese hamster ovary (CHO) cells and preparation method thereof
  • Protein-free culture medium for culturing microencapsulated recombinant Chinese hamster ovary (CHO) cells and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] 1. Prepare protein-free medium for microencapsulated recombinant CHO cells:

[0035] Culture medium of the present invention is made up of two parts: DMEM / F12 culture medium (purchased from Sigma Company, composition sees Table 1) and following composition:

[0036]

[0037] Table 1 DMEM / F12 medium composition

[0038]

[0039] All the above substances are chemical reagents of analytical grade, dissolved in ultrapure water with a final volume of 90%, and after fully stirring for half an hour, dilute to the required final volume and keep for 10 minutes.

[0040] Sterile filtration is carried out according to the standard operation of flat filter.

[0041] 2. Preparation of microencapsulated recombinant CHO cells:

[0042] Reference literature (document 4. Ma X J, 1994. Generation of alginate-poly-Lysine alginate (APA) biomicrocapsules: the relationship between the membrane strength and the reaction conditions. Art Cells Blood Subs and Immob Biotech, 22 (1): 43- ...

Embodiment 2

[0045] 1. Prepare protein-free medium for microencapsulated recombinant CHO cells:

[0046] Culture medium of the present invention is made up of two parts: DMEM / F12 culture medium (purchased from Sigma Company, composition sees Table 1) and following composition:

[0047]

[0048] 2. Preparation of microencapsulated recombinant CHO cells (same as Example 1):

[0049] Microencapsulate cells at 37 °C, 5% CO 2 In an incubator, culture in the medium prepared in the above step 1 of this embodiment for 7 days to measure the expression level of DSPA, and compare it with the control example.

Embodiment 3

[0051] 1. Prepare protein-free medium for microencapsulated recombinant CHO cells:

[0052] Culture medium of the present invention is made up of two parts: DMEM / F12 culture medium (purchased from Sigma Company, composition sees Table 1) and following composition:

[0053]

[0054] 2, the preparation of empty microcapsules (do not add cell, all the other are with embodiment 1):

[0055] microcapsules at 37 °C, 5% CO 2 In an incubator, culture the microcapsules for 7 days in the medium prepared in step 1 of this embodiment to observe the stability of the microcapsules.

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Abstract

The invention discloses a protein-free culture medium for culturing microencapsulated recombinant Chinese hamster ovary (CHO) cells and a preparation method thereof. By using DMEM (dulbecco's modified eagle medium)/F12 as a basal culture medium, micromolecule substance iron salt with definite component, zinc salt, sodium selenite, sodium acetate, glutamine and glucose are added according to the characteristics of the microencapsulated cell culture. The culture medium does not influence the stability of the microcapsules, does not have insulin or transferrin in the traditional serum-free culture medium, and lowers the downstream technique and cost; and the product expression level is approximate to the culture level of the expensive specific culture medium, but the price is only one-tenth of the expensive specific culture medium. Thus, the protein-free culture medium can be used for microencapsulated recombinant CHO cells culture and protein expression, and greatly lowers the culture cost.

Description

technical field [0001] The present invention relates to the field of cell culture. More specifically, the present invention relates to a protein-free medium for culture of microencapsulated recombinant Chinese Hamster Ovary (CHO) cells and a corresponding culture method. Background technique [0002] Microencapsulated cell culture was first proposed by Chang TMS (Document 1. Chang TMS, 1964. Semipermeable microcapsules. Science 146, 524-525) in 1964. It is a widely used culture technology and has been used for large-scale cell culture and recombination. fields of protein production. Microcapsules can increase the stress tolerance of cells to the physical and chemical environment, and realize high-density culture of cells (document 2, Rokstad AM, 2002. Microencapsulation of cells producing therapeutic proteins: Optimizing cell growth and secretion. Cell Transplant 11 :313-324), significantly improved product expression (document 3.Zhang Y, 2008.Optimization of microencapsul...

Claims

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Application Information

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IPC IPC(8): C12N5/10C12N11/10C12N11/04
Inventor 马小军王雨张英李娜陈立于炜婷李珅
Owner DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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