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Serum-free culture medium of human embryo kidney (HEK) 293 cell

A serum-free medium and cell culture technology, applied in the field of medium, can solve the problem of high price of serum-free medium

Inactive Publication Date: 2007-02-07
EAST CHINA UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] At present, there are serum-free media suitable for HEK293 cell culture: 293-SFMII (Invitrogen), Ex-Cell 525 (JRH), IS293-SF (Irvine Scientific), Pro293s-CDM (Cambrex), CD293 Medium, Freestyle TM 293 and Expression Medium (Gibco), etc., but these serum-free media are more expensive

Method used

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  • Serum-free culture medium of human embryo kidney (HEK) 293 cell

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Embodiment 1

[0100] The said preparation of the serum-free medium that is used for human embryonic kidney (HEK) 293 cell culture of the present invention:

[0101] Classify and dissolve the following raw materials according to their respective solubility characteristics (according to the operation steps provided by the supplier), then mix the resulting solution at 20-30°C so that the contents of each component are listed below, and adjust the pH value of the resulting mixture to 7.2 , the target object is obtained after constant volume.

[0102] product name

[0103] product name

[0104] product name

[0105] product name

Embodiment 2

[0107] Another serum-free medium for culturing human embryonic kidney (HEK) 293 cells was prepared according to the method described in Example 1: its composition and content are as follows:

[0108] L-alanine 30mg / L

[0109] L-Arginine 300mg / L

[0110] L-glutamic acid 150mg / L

[0111] L-Asparagine 200mg / L

[0112] L-Aspartic Acid 300mg / L

[0113] L-cysteine ​​300mg / L

[0114] L-cystine 600mg / L

[0115] L-Glycine 50mg / L

[0116] L-histidine 700mg / L

[0117] L-Isoleucine 700mg / L

[0118] L-leucine 1200mgL

[0119] L-Lysine 1500mg / L

[0120] L-methionine 400mg / L

[0121] L-phenylalanine 500mg / L

[0122] L-proline 50mg / L

[0123] L-Threonine 600mg / L

[0124] L-serine 200mg / L

[0125] L-Tryptophan 200mg / L

[0126] L-tyrosine 800mg / L

[0127] L-Valine 700mg / L

[0128] L-Glutamine 9000mg / L

[0129] Vitamin section:

[0130] Biotin 0.01mg / L

[0131] Choline chloride 20mg / L

[0132] Calcium pantothenate 10mg / L

[0133] Folic acid 5mg / L

[0134] Inositol 20mg / L ...

Embodiment 3

[0172] HEK293 cells adapted to serum-free suspension culture were cultured in a 125ml square flask at a seeding density of 3×10 5 cells / ml, the filling volume is 20ml, and then the square flask with the seeded cells is placed on a shaker (model: TZ-03, SHENERGYBIOCOLOR) for culture (the shaker is placed at 36.5 ° C, 5% CO 2 incubator), the rotation speed was 120r / min, cultured for 5 days, and the maximum cell density reached 3.8×10 6 cells / ml. The result is as figure 1 . The serum-free medium used in this example is prepared from Example 1.

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Abstract

the invention discloses a non-serum substratum for cell of HEK 293, which is composed of transferring, insulin, Primatone, lipoid compound, amino acid, inorganic salt and vitamin based on basic substratum.

Description

technical field [0001] The invention relates to a medium for culturing human embryonic kidney (HEK) 293 cells, in particular to a serum-free medium for culturing HEK293 cells. Background technique [0002] Human embryonic kidney (HEK) 293 cells were first established by Graham et al. in 1977 and are ideal hosts for the production of recombinant adenoviruses. [0003] The traditional culture medium for HEK293 cells is a medium with serum. However, with the increasing awareness that animal serum may contaminate the production process and even the final product, the use of serum-containing media has been limited. Serum-free medium has attracted more and more attention because of its advantages of non-source pollution, small batch-to-batch variability, and favorable separation and purification of expression products. [0004] At present, there are serum-free media suitable for HEK293 cell culture: 293-SFMII (Invitrogen), Ex-Cell 525 (JRH), IS293-SF (Irvine Scientific), Pro293s...

Claims

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Application Information

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IPC IPC(8): C12N5/08C12N5/073
Inventor 易小萍陈春艳张元兴孙祥明
Owner EAST CHINA UNIV OF SCI & TECH
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