Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

A kind of serum-free medium and application for large-scale cultivation of cho cells

A technology of serum-free culture medium and culture medium, which is applied in the cultivation process, animal cells, tissue culture, etc., to achieve the effect of simple ingredients, low cost, and improvement of clumping phenomenon

Active Publication Date: 2022-05-27
SHANGHAI OPM BIOSCI CO LTD
View PDF6 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] In order to solve the deficiencies of the existing CHO cell suspension culture, the invention provides a serum-free medium for cultivating CHO cells and its application

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • A kind of serum-free medium and application for large-scale cultivation of cho cells
  • A kind of serum-free medium and application for large-scale cultivation of cho cells
  • A kind of serum-free medium and application for large-scale cultivation of cho cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0051] The preparation of embodiment 1 culture medium and cell line

[0052] Basal medium: DMEM / F12 is a commercial basal medium commonly used in cell culture. The medium is composed of glucose, amino acids, vitamins, inorganic salts and other substances. Available from Sigma, GIBCO, etc.

[0053] On the basis of DMEM / F12, three kinds of serum-free medium for culturing CHO cells of the present invention with different concentration ratios were prepared.

[0054] Component, concentration mg / L Experimental example 1 Experimental example 2 Experimental example 3 AEO-9 100 200 150 cholesterol linoleate 5 10 7 Vitamin C 5 10 8 insulin 2 5 3 Transferrin 5 7 6 Ethanolamine 2 5 3 Lecithin 3 6 5 Choline Sulfate 10 40 25

[0055] Trace elements are all added in the above-mentioned experimental examples 1-3, and the final concentrations of each component of the trace elements are respectively: zinc sulf...

Embodiment 2

[0061] Example 2 Batch suspension culture of rCHO-GS cells in a 5-liter stirred tank bioreactor

[0062] Cell culture: rCHO-GS cells were suspended and cultured in the medium of Experimental Example 1, Experimental Example 2, Experimental Example 3, Comparative Example 2, and Comparative Example 3 in a 250-m1 stirring flask, at a rate of 3 × 10. 5 cells / ml were inoculated into a 5L stirred tank bioreactor, and the above-mentioned medium was added to the set working volume of 5L. The temperature was set to 37°C, the dissolved oxygen concentration was controlled at 30-50%, the pH was controlled at 7.1-7.2, and the stirring speed was set at 60-70r / min. Samples were taken every day, and the cells were counted with a hemocytometer, and each sample was counted 3 times. , take the average value, and determine the viability of cells by trypan blue exclusion method. The whole culture process lasted 7d. The different results using the above 5 media are shown in Tables 2, 3 and figur...

Embodiment 3

[0067] Example 3 Suspension culture of wild-type CHO cells

[0068] Experimental Example 3 and Comparative Example 3 in Example 2 were repeated, except that rCHO-GS cells were replaced with wild-type CHO cells, ie, CHO-K1ATCC Cat.NO.CC1-61. The experimental results showed that, using the medium of Experimental Example 3, the wild-type CHO cells reached the maximum cell density of 79.5×10 on the sixth day. 6 / mL, Comparative Example 3 also reached the maximum cell density on the sixth day, but only 58.1 × 10 6 / mL. After 7 days of culture, using the medium of Experimental Example 3, the survival rate of wild-type CHO cells was always above 90%. However, using the medium of Comparative Example 3, the survival rate of wild-type CHO cells was only 75.9% on day 7 of culture. It can be seen that using the serum-free medium of the present invention to cultivate rCHO-GS or wild-type CHO cells can achieve better results. Compared with the commercially available CHO serum-free medium ...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention relates to a serum-free medium for large-scale cultivation of CHO cells and its application. The culture medium contains basal medium and supplements, the basal medium is DMEM / F12, and the supplements include AEO‑9, cholesterol linolenate, vitamin C, insulin, transferrin, ethanolamine, lecithin , choline chloride and trace elements. The medium can be used for industrialized large-scale cultivation of CHO cells, and supports better growth, higher survival rate and less agglomeration of CHO cells.

Description

technical field [0001] The present invention relates to the field of cell culture, more particularly, to a serum-free medium for culturing CHO cells and application thereof, the medium can be used for industrialized large-scale culture of CHO cells, and supports better growth of CHO cells in suspension culture , higher survival rate and less clumping. Background technique [0002] CHO cells (Chinese hamster ovary cells, Chinese hamster ovary cells) were isolated from an adult female hamster ovary by Dr. Theodore T. Puck of the University of Colorado in 1957. They are epithelial adherent cells and are widely used in bioengineering. . The cells are immortal and can be passaged for more than 100 generations, and are widely used cells in bioengineering. In addition, CHO cells have another advantage in the use of genetic engineering. The cells belong to fibroblasts, which are non-secreting cells. They rarely secrete CHO endogenous proteins, so they are very beneficial for the s...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/071C12N5/02
CPCC12N5/0682C12N2500/90C12N2500/50C12N2500/36C12N2500/38C12N2500/25C12N2500/46C12N2500/30C12N2500/22C12N2500/20C12N2500/12C12N2500/14C12N2500/32C12N2500/24C12N2500/16C12N2500/34C12N2500/40
Inventor 肖志华邓鸿云
Owner SHANGHAI OPM BIOSCI CO LTD
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com