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Serum-free protein-free culture medium supporting CHO (Chinese Hamster Ovary Cell) high density suspension culture

A protein-free medium and suspension culture technology, applied in tissue culture, microorganisms, biochemical equipment and methods, etc., can solve the problems of being easily affected by physical and chemical factors, pollution separation and purification, and high cost, and achieve the goal of being conducive to separation and purification. , the risk of contamination is small, the effect of chemical composition is clear

Active Publication Date: 2014-10-01
SUNSHINE GUOJIAN PHARMA (SHANGHAI) CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, serum-free medium usually contains proteins and lipids that replace serum, such as transferrin, lipid additives and insulin, etc., there are still certain contamination risks and barriers to separation and purification, and the cost is relatively high
Moreover, generally a serum-free medium is only suitable for the cultivation of one type of cells, and is easily affected by physical and chemical factors.

Method used

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  • Serum-free protein-free culture medium supporting CHO (Chinese Hamster Ovary Cell) high density suspension culture
  • Serum-free protein-free culture medium supporting CHO (Chinese Hamster Ovary Cell) high density suspension culture
  • Serum-free protein-free culture medium supporting CHO (Chinese Hamster Ovary Cell) high density suspension culture

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] CHO cell serum-free protein-free medium dry powder components and dosage of the present invention are as follows:

[0040]

[0041]

[0042]

[0043]

[0044] Medium dry powder configuration method (1L):

[0045] Add 1L of dry powder into 900ml of ultrapure water at a temperature of about 35°C. After stirring for half an hour, add a certain amount of sodium hydroxide to aid dissolution, then add concentrated hydrochloric acid and sodium bicarbonate to adjust the pH to 6.9-7.4. Filter with 0.2um negative pressure, aliquot into 500ml glass bottles and store aseptically in the dark.

[0046] The physical and chemical property parameters, detection methods and detection results of CHO cell serum-free protein-free medium culture that the present invention obtains are as shown in Table 1:

[0047] Table 1

[0048] Test items

[0049] From the test results, the configured liquid culture medium is qualified and can be used for CHO large-scale suspension...

Embodiment 2

[0051] Insulin and transferrin were added on the basis of the serum-free and protein-free medium in Example 1, and the components and consumption of the medium dry powder were as follows:

[0052]

[0053]

[0054]

Embodiment 3

[0056] On the basis of the serum-free and protein-free medium of Example 1, the ultra-filtered hydrolyzate (molecular weight less than 1000 Daltons) of plant origin is added, and the components and dosage of the medium dry powder are as follows:

[0057]

[0058]

[0059]

[0060]

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Abstract

The invention provides a serum-free protein-free culture medium supporting CHO (Chinese Hamster Ovary Cell) high density suspension culture. The culture medium comprises amino acids, microelements, inorganic salts, vitamins, carbohydrates and other organic molecules, wherein culture medium contains steroid hormone but does not contain transferrin or insulin. The culture medium provided by the invention is definite in chemical component, free of animal resource, small in pollution risk, applicable to growth of various different CHO cell strains, good in cell culture effect and beneficial for downstream separation and purification.

Description

technical field [0001] The present invention relates to the field of cell culture. More specifically, the present invention relates to culture media for CHO cell culture. Background technique [0002] Cell culture technology is to achieve cell culture in vitro by simulating the growth environment of cells in vivo. Culture medium provides nutrients for in vitro culture of cells and promotes cell growth and proliferation. The development of cell culture medium has gone through chicken embryo juice, natural medium, synthetic medium, and currently commonly used low serum medium, serum-free medium, protein-free medium and medium with clear chemical composition. [0003] Usually, the growth of animal cells depends on the presence of serum. In ordinary medium, most cells will not be able to proliferate without adding serum. However, the main disadvantages of using serum are potential sources of contamination, high price, and large batch-to-batch variance, which brings a lot of ...

Claims

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Application Information

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IPC IPC(8): C12N5/071C12N5/02
Inventor 胡辉李军翁志兵
Owner SUNSHINE GUOJIAN PHARMA (SHANGHAI) CO LTD
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