CH150 medium｜CHO cells｜Single Cell Cloning｜Chemically Defined Medium

CHO Chemically Defined Media

① Adherent Culture

・CH100 medium

② Cloning Assay

・CH150 medium

③ Suspension Culture

・CH200 medium

・CH300 medium

・CH300AZ medium＊

・CH400 medium

・CH400AZ medium＊

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

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④ Fed-Batch Culture

・Gxpress CHO Feed medium

⑤ Anti-Clumping Reagent

・Scattering reagent CHO

Single Cell Cloning Assay
CH150 medium (CD medium)
For CHO cells "CHO-K1, CHO-S, DG44, DXB11, etc."

CHO cells｜Single cell cloning assay｜Chemically defined & Serum-free medium

CH150 medium is a chemically defined medium / serum-free medium optimized for the single-cell cloning of Chinese hamster ovary (CHO) cells, such as CHO-K1, CHO-S, DG44, and DXB11 cells. The medium contains no proteins, hydrolysates, or components of animal and plant composition. All components have a known chemical structure.
Single-cell cloning in CHO 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. CH150 medium has enabled a single-cell cloninng for CHO cells.

Protocol

・CH150 medium protocol

Figure 1）Single cell cloning assay of CHO cells using CH150 medium.

Suspension CHO cells were cultured with 96-well plate under the following conditions.
CH150 medium is a medium that can lead to single cell growth for CHO-S cells and CHO-K1 cells.
＜ Culture conditions ＞
(1) Seeding cell density：1 cell/well.
(2) Static culture：37℃, 5% CO2.
(3) Culture volume：200 μL with L-glutamine.

Figure 2) Growth assay of CHO-K1 cells cloned by CH150 medium.

The growth assay was performed using CH400AZ medium on clonal CHO-K1 cells obtained by cloning assay with CH150 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：2 x 10e5 cells/mL.
(2) Shaker culture：125 rpm, 37℃, 8% CO2.
(3) Culture volume：30 mL with L-alanyl-L-glutamine.

Figure 3) Growth assay of CHO-S cells cloned by CH150 medium.

The growth assay was performed using CH400AZ medium on clonal CHO-S cells obtained by cloning assay with CH150 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：2 x 10e5 cells/mL.
(2) Shaker culture：125 rpm, 37℃, 8% CO2.
(3) Culture volume：30 mL with L-alanyl-L-glutamine.