HE300AZ medium｜293 cells｜Suspension Culture｜Chemically Defined Medium

293 Chemically Defined Media

① Adherent Culture

・HE100 medium

② Cloning Assay

・HE150 medium

③ Suspension Culture

・HE200 medium

・HE300 medium

・HE300AZ medium＊

・HE400 medium

・HE400AZ medium＊

＊Ready-to-use products with L-alanyl-L-glutamine

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④ Fed-Batch Culture

・Gxpress 293 Feed medium

Suspension Culture (Expression & Production)
HE300AZ medium (CD medium), liquid or powder
For 293 cells "Expi293F, 293-F, HEK293, 293T, VPC, etc."
*A ready-to-use product that contains L-alanyl-L-glutamine.

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

　HE300AZ 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, 293T, VPC cells. The medium contains no proteins, hydrolysates, or human/animal-derived components. All components have a known chemical structure.

HE300AZ 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.

Protocol

・HE300AZ medium protocol

Figure 1) Growth assay of Expi293F cells in HE300/HE300AZ medium.

Suspension 293 cells were cultured with disposable Erlenmeyer flasks under the following conditions.
HE300/HE300AZ medium leads to the high performance similar to Thermo FreeStyle 293 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 HE300/HE300AZ medium.

Suspension 293 cells were cultured with disposable Erlenmeyer flasks under the following conditions.
HE300/HE300AZ medium leads to the high performance 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 HE300/HE300AZ medium.

HE300/HE300AZ medium exhibits the equal or 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 HE300/HE300AZ medium.

HE300/HE300AZ medium exhibits the equal or 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) Growth assay of 293T cells or HEK293 cells in HE300/HE300AZ medium.

Suspension 293 cells were cultured with disposable Erlenmeyer flasks under the following conditions.
HE300/HE300AZ medium leads to the high performance similar to Thermo FreeStyle 293 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.