HE200 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)
HE200 medium (CD medium), liquid or powder
For 293 cells "Expi293F, 293-F, HEK293, 293T, VPC, etc."

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

　HE200 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, or VPC cells. The medium contains no proteins, hydrolysates, or human/animal-derived components. All components have a known chemical structure.

HE200 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

・HE200 medium protocol

Figure 1) Growth assay of Expi293F cells in HE200 medium.

Suspension 293 cells were cultured with disposable Erlenmeyer flasks under the following conditions.
HE200 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 HE200 medium.

Suspension 293 cells were cultured with disposable Erlenmeyer flasks under the following conditions.
HE200 medium leads to the high performance similar to Thermo FreeStyle 293 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 HE200 medium.

HE200 medium exhibits the equal 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 HE200 medium.

HE200 medium exhibits the equal or higher performance in gene expression compared to Thermo FreeStyle 293 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 HE200 medium.

Suspension 293 cells were cultured with disposable Erlenmeyer flasks under the following conditions.
HE200 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.