293 Chemically Defined Media
Single Cell Cloning Assay
HE150 medium (CD medium)
For 293 cells "Expi293F, 293-F, HEK293, 293T, VPC, etc."
HE150 medium is a chemically defined medium / serum-free medium optimized for the single-cell growth of 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.
Single-cell cloning in HEK293 cells is an important step in producing homogenous recombinant protein and allows homogeneity of a selected cell line. In general, serum-free medium results in very low cell cloning efficientry, and also typically fails to establish a stable cell clone producing recombinant protein. HE150 medium has enabled a single-cell cloninng for 293 cells.
Figure 1）Single cell cloning assay of 293 cells using HE150 medium.
Suspension 293 cells were cultured with 96-well plate under the following conditions.
HE150 medium is a medium that can lead to single cell growth for Expi293F cells and 293-F cells.
＜ Culture conditions ＞
(1) Seeding cell density：1 cell/well.
(2) Static culture：37℃, 5% CO2.
(3) Culture volume：200 μL with L-alanyl-L-glutamine.
The growth assay was performed using HE400AZ medium on clonal Expi293F cells obtained by cloning assay with HE150 medium.
Clonal cells had the ability to grow to higher cell densities compared to the cells before cloning assay.
＜ 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）Single cell cloning assay of 293 cells using HE150 medium.
Suspension 293 cells were cultured with 96-well plate under the following conditions.
HE150 medium is a medium that can lead to single cell growth for 293T cells and HEK293 cells.
These cells are suspended with adaptation in HE400AZ medium.
＜ Culture conditions ＞
(1) Seeding cell density：1 cell/well.
(2) Static culture：37℃, 5% CO2.
(3) Culture volume：200 μL with L-alanyl-L-glutamine.
Product Lineup
Gmep Incorporated
Hyakunen Park, Kurume, Fukuoka, Japan