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Non-serum culture medium adapted for large-scale Chinese hamster ovary cells culture

A technology of large-scale culture and medium, applied in the field of cell culture, it can solve the problems of low cell density and protein expression, lack of it, etc., and achieve the effect of good culture effect.

Inactive Publication Date: 2008-02-06
SUNSHINE GUOJIAN PHARMA (SHANGHAI) CO LTD
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  • Claims
  • Application Information

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Problems solved by technology

[0007] In summary, although scientists from various countries have developed many serum-free media, most of them are only suitable for one or a type of cell growth, especially lack of suitable for large-scale CHO cell culture, and the culture effect is similar to serum-free media. Medium equivalent or better medium, and for commercially available serum-free medium such as medium Excell325 that can be used for large-scale CHO cell culture PF There are also problems such as low cell density and protein expression during use

Method used

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  • Non-serum culture medium adapted for large-scale Chinese hamster ovary cells culture
  • Non-serum culture medium adapted for large-scale Chinese hamster ovary cells culture
  • Non-serum culture medium adapted for large-scale Chinese hamster ovary cells culture

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Experimental program
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Effect test

Embodiment 1

[0072] Preparation and CHO cell culture of embodiment 1 medium

[0073] (1) Culture medium formula: wherein the basal medium DMEM / F12 medium (purchased from Sigma Company), on the basis of the medium, add the following substances:

[0074] Vitamin C 10.5mg / L;

[0075] Transferrin 6.2mg / L;

[0076] Ethanolamine 3.1mg / L;

[0077] Selenite 0.025mg / L;

[0078] Sodium hydroxybutyrate 0.85mg / L, based on the total volume of CHO cell culture medium.

[0079] Trace elements:

[0080] AlCl 3 ·6H 2 O 1.2mg / L

[0081] AgNO 3 0.03mg / L

[0082] CoCl 2 ·6H 2 O 0.21mg / L

[0083] CuSO 4 ·5H 2 O 0.0027mg / L

[0084] KBr 0.00061mg / L

[0085] FeSO 4 ·7H 2 O 0.72 mg / L

[0086] Na 2 SiO 3 0.005~0.045mg / L

[0087] LiCl 0.00083mg / L

[0088] NiCl 2 ·6H 2 O 0.042mg / L

[0089] SnCl 2 2H 2 O 2.12mg / L

[0090] ZnSO 4 ·7H 2 O 1.58mg / L

[0091] Vitamins:

[0092] Vitamin B6 0.18mg / L

[0093] Vitamin B12 0.38mg / L

[0094] Biotin 0.12 mg / L

[0095] Folic ac...

Embodiment 2

[0113] Preparation and CHO cell culture of embodiment 2 medium

[0114] (1) Culture medium formula: wherein the basal medium DMEM / F12 medium (purchased from Sigma Company), on the basis of the medium, add the following substances:

[0115] Vitamin C 10.5mg / L;

[0116] Transferrin 6.2mg / L;

[0117] Ethanolamine 3.1mg / L;

[0118] Selenite 0.025mg / L;

[0119] Sodium hydroxybutyrate 0.85mg / L, based on the total volume of CHO cell culture medium.

[0120] Trace elements:

[0121] AlCl 3 ·6H 2 O 1.2mg / L

[0122] AgNO 3 0.03mg / L

[0123] CoCl 2 ·6H 2 O 0.21mg / L

[0124] CuSO 4 ·5H 2 O 0.0027mg / L

[0125] KBr 0.00061mg / L

[0126] FeSO 4 ·7H 2 O 0.72mg / L

[0127] Na 2 SiO 3 0.005~0.045mg / L

[0128] LiCl 0.00083mg / L

[0129] NiCl 2 ·6H 2 O 0.042mg / L

[0130] SnCl 2 2H 2 O 2.12mg / L

[0131] ZnSO 4 ·7H 2 O 1.58mg / L

[0132] Vitamins:

[0133] Vitamin B6 0.18mg / L

[0134] Vitamin B12 0.38mg / L

[0135] Biotin 0.12mg / L

[0136] Folic acid 3...

Embodiment 3

[0143] The preparation of embodiment 3 medium and the large-scale culture of CHO cell

[0144] (1) Culture medium formula: wherein the basal medium DMEM / F12 medium (purchased from Sigma Company), on the basis of the medium, add the following substances:

[0145] Vitamin C 10.5mg / L;

[0146] Transferrin 6.2mg / L;

[0147] Ethanolamine 3.1mg / L;

[0148] Selenite 0.025mg / L;

[0149] Sodium hydroxybutyrate 0.85mg / L, based on the total volume of CHO cell culture medium.

[0150] Trace elements:

[0151] AlCl 3 ·6H 2 O 1.2mg / L

[0152] AgNO 3 0.03mg / L

[0153] CoCl 2 ·6H 2 O 0.21mg / L

[0154] CuSO 4 ·5H 2 O 0.0027mg / L

[0155] KBr 0.00061mg / L

[0156] FeSO 4 ·7H 2 O 0.72mg / L

[0157] Na 2 SiO 3 0.005~0.045mg / L

[0158] LiCl 0.00083mg / L

[0159] NiCl 2 ·6H 2 O 0.042mg / L

[0160] SnCl 2 2H 2 O 2.12mg / L

[0161] ZnSO 4 ·7H 2 O 1.58mg / L

[0162] Vitamins:

[0163] Vitamin B6 0.18mg / L

[0164] Vitamin B12 0.38mg / L

[0165] Biotin 0.12m...

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Abstract

The present invention provides a serum free medium used for cultivating CHO cellular extensively, and discloses the prescription of the culture medium. DMEM / F12 is used as a base culture medium and is added with the components such as vitamin C, transferring, neovaricaine, hydroxyl sodium butyrate and so on. The present invention has the advantages that no animal blood serum is contained and the content of albumen is low; the cultivating effect of the CHO cellular is good, and the CHO cellular can obtain the same effect with the culture medium containing blood serum.

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] Chinese hamster ovary cells—CHO cells (Chinese hamster ovary cells) are currently widely used to produce various genetically engineered protein products. The medium suitable for CHO cell culture is divided according to its source and the degree of clarity of its components. Its development can be roughly divided into three stages: the natural medium stage (direct use of certain tissue clots, biological fluids and Tissue extract, etc. as the culture medium of cells), synthetic medium stage (such as MEM, DMEM, RPMI1640, F12, etc.) and serum-free medium stage. Synthetic medium usually needs to add some natural body fluid supplements such as serum (commonly used such as calf serum, fetal calf serum, etc., generally at 5-15%) to maintain cell growth for a long time. The main func...

Claims

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Application Information

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IPC IPC(8): C12N5/06C12N5/071
Inventor 寇庚王皓钱卫珠
Owner SUNSHINE GUOJIAN PHARMA (SHANGHAI) CO LTD
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