Serum-free medium for culturing insect cells in full suspension mode and preparation method and application thereof

A technology of serum-free medium and insect cells, applied in biochemical equipment and methods, culture process, tissue culture, etc., can solve the problem of unsatisfactory cell growth

Active Publication Date: 2016-08-17
NORTHWEST UNIVERSITY FOR NATIONALITIES
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Serum-free medium BFPM401 is a domestically produced medium suitable for high-density mammalian cells. At present, there...

Method used

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  • Serum-free medium for culturing insect cells in full suspension mode and preparation method and application thereof
  • Serum-free medium for culturing insect cells in full suspension mode and preparation method and application thereof
  • Serum-free medium for culturing insect cells in full suspension mode and preparation method and application thereof

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Embodiment 1

[0049] 1 Materials and Equipment

[0050] 1.1 Materials

[0051] Insect cells: Sf-21, Hi-five (provided by Gansu Animal Cell Engineering Technology Research Center);

[0052] Medium: Grace’s Insect Medium dry powder (Gibco), serum-free medium BFPM401 dry powder (Lanzhou Bailing Biology);

[0053] Control medium HyQ SFX-Insect SFM (Hyclone, liquid, used directly), glucose (Sinopharm Group), hydrolyzed milk protein (HyClone), Pluronic F-68 (Gibco), yeast powder (Organotechnie), etc.

[0054] 1.2 Equipment

[0055] Inverted phase contrast microscope (CK40-32PH, Olympus), cell counter (CASY TT, INNOVATIS), electronic balance (AR2140, Shanghai Ohaus), shaker (2102C, Shanghai Zhicheng), pH meter (MP120 -BLE type, Mettler) etc.

[0056] 2 methods

[0057] 2.1 Dosing

[0058] Preparation of yeast powder concentrate with a concentration of 50 times: Weigh 200g of yeast powder and dissolve it in 500ml of water for injection, stir repeatedly and put it in a refrigerator at 2-8°C, t...

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Abstract

The invention provides a serum-free medium for culturing insect cells in a full suspension mode. The serum-free medium is prepared from a Grace's Insect Medium, a BFPM410 medium, 2-4 g/L of glucose, 1-3 g/L of lactalbumin hydrolysate, 1-3 g/L of PF68 and 2-5 g/L of yeast powder. The invention further provides a preparation method and application of the serum-free medium. As shown in experiments, the serum-free medium can be well used for culturing insect cells in the full suspension mode, the maximum cell proliferation concentration is 42.5*10<5>/mL when the serum-free medium is used for culturing Sf-21 cells, and the maximum cell proliferation concentration is 45.4*10<5>/mL when the serum-free medium is used for culturing Hi-five cells.

Description

technical field [0001] The invention belongs to the technical field of biological products, and in particular relates to a serum-free culture medium for fully suspension culturing insect cells. Background technique [0002] Baculovirus Expression Vector System (BEVS) is a eukaryotic expression system that uses baculovirus as a vector to express foreign proteins on insect cells. Because the insect baculovirus system can produce heterologous proteins at a high level and ensure the biological activity of heterologous proteins, it has been applied in medical diagnosis and vaccine production. The vaccines produced by the expression system of insect baculovirus that are currently on the market include classical swine fever vaccine, porcine circovirus type 2 vaccine, cervical cancer vaccine and therapeutic prostate cancer vaccine. In addition, in 2013, the US FDA approved Flublok for the prevention of seasonal influenza in people aged 19 to 49. Flublok also became the first influe...

Claims

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Application Information

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IPC IPC(8): C12N5/07C12N5/02
CPCC12N5/0601C12N2500/30C12N2500/34C12N2500/74C12N2500/90C12N2501/998
Inventor 马忠仁乔自林马桂兰冯玉萍马伟王家敏马祺
Owner NORTHWEST UNIVERSITY FOR NATIONALITIES
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