HOME | HE400AZ meidium|293 cells|Suspension Culture|Chemically Defined Medium

293 Chemically Defined Media

① Adherent Culture
 
② Cloning Assay
③ Suspension Culture
*Ready-to-use products with L-alanyl-L-glutamine
④ Fed-Batch Culture

Suspension Culture (Expression & Production)
HE400AZ medium (CD medium), liquid or powder
For 293 cells "Expi293F, 293-F, HEK293, 293T, etc."
*A ready-to-use product that contains L-alanyl-L-glutamine.

293 cells|Suspension culture|Chemically defined & Serum-free medium

 HE400AZ medium is a chemically defined medium / serum-free medium optimized for the high production of recombinant proteins in human embryonic kidney (HEK) 293 cells, such as Expi293F, 293-F, HEK293, or 293T cells. The medium contains no proteins, hydrolysates, or components of animal and plant composition. All components have a known chemical structure.

   HE400AZ medium has the advantages of lot-to-lot consistency and the ease of downstream purification after expression of recombinant proteins. The medium supports small- and large-scale protein expression in stably or transiently transfected 293 cells.

Figure 1) Growth assay of Expi293F cells in HE400/HE400AZ medium.


Suspension 293 cells were cultured with disposable Erlenmeyer flasks under the following conditions.
HE400/HE400AZ medium leads to the high performance similar to Thermo Expi293 Expression medium in cell growth for Expi293F cells.
< Culture conditions >
(1) Seeding cell density:3 x 10e5 cells/mL.
(2) Shaker culture:125 rpm, 37℃, 8% CO2.
(3) Culture volume:30 mL with L-alanyl-L-glutamine.


Figure 2) Growth assay of 293-F cells in HE400/HE400AZ medium.


Suspension 293 cells were cultured with disposable Erlenmeyer flasks under the following conditions.
HE400/HE400AZ medium leads to the high performance similar to Thermo Expi293 Expression medium in cell growth for 293-F cells.
< Culture conditions >
(1) Seeding cell density:3 x 10e5 cells/mL.
(2) Shaker culture:125 rpm, 37℃, 8% CO2.
(3) Culture volume:30 mL with L-alanyl-L-glutamine.


Figure 3) Transient transfection assay of Expi293F cells with HE400/HE400AZ medium.

HE400/HE400AZ medium exhibits the higher performance in human IgG production compared to Thermo Expi293 Expression medium without inhibiting the transfection assay.
< Culture conditions >
(1) Seeding cell density:2.94 x 10e6 cells/mL, >95% viability.
(2) Shaker culture:125 rpm, 37℃, 8% CO2.
(3) Transfection reagent:Gxpress 293 TF Reagent (GMEP Inc.).


Figure 4) Transient transfection assay of Expi293F cells with HE400/HE400AZ medium.

HE400/HE400AZ medium exhibits the higher performance in gene expression compared to Thermo Expi293 Expression medium without inhibiting the transfection assay.
< Culture conditions >
(1) Seeding cell density:2.94 x 10e6 cells/mL, >95% viability.
(2) Shaker culture:125 rpm, 37℃, 8% CO2.
(3) Transfection reagent:Gxpress 293 TF Reagent (GMEP Inc.).


Figure 5) HE400/HE400AZ medium shows supperior performance in the cell culture of Expi293F cells.

Cryopreserved Expi293F cells were thawed using Thermo Expi293 Expression medium or HE400/HE400AZ medium, and subcultured in disposable Erlenmeyer flasks under the fillowing conditions.
Next, the subculture was repeated with each medium to allow the cells to recover cryopreservation. Expi293F cells subcultured were subjected to growth assay using both Expi293 Expression medium and HE400/HE400AZ medium.
Similar to the cells passaged in Expi293 Expression medium, Expi293F cells subcultured with HE400/HE400AZ medium can exert the high performance in cell culture.
< Culture conditions >
(1) Seeding cell density:3 x 10e5 cells/mL.
(2) Shaker culture:125 rpm, 37℃, 8% CO2.
(3) Culture volume:30 mL with L-alanyl-L-glutamine.

Figure 6) Growth assay of 293T cells or HEK293 cells in HE400/HE400AZ medium.


Suspension 293 cells were cultured with disposable Erlenmeyer flasks under the following conditions.
HE400/HE400AZ medium leads to the high performance similar to Thermo Expi293 Expression medium in cell growth for 293T cells or HEK293 cells.
These cells cultured in DMEM with 10% FBS are suspended with adaptation in HE400AZ medium.
< Culture conditions >
(1) Seeding cell density:3 x 10e5 cells/mL.
(2) Shaker culture:125 rpm, 37℃, 8% CO2.
(3) Culture volume:30 mL with L-alanyl-L-glutamine.


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