title: Tetrahydrofolate increases suspension growth of dihydrofolate reductase-deficient chinese hamster ovary DG44 cells in chemically defined media.
abstract: Adaptation of dihydrofolate reductase (DHFR)-deficient Chinese hamster ovary (CHO) DG44 cells to chemically defined suspension culture conditions is a time-consuming and labor-intensive process because nonadapted DHFR-deficient CHO DG44 cells normally show poor growth in chemically defined medium (CDM). We examined the effects of folate derivatives, ribonucleotides, and nucleobases on the growth of suspension-adapted DHFR-deficient CHO DG44 cells in CDM. Among the tested additives, tetrahydrofolate (THF) was identified as an effective component for increasing cell growth. THF supplementation in the range of 0.2-359 μM enhanced cell growth in in-house CDM. Addition of 3.6 μM THF to in-house CDM resulted in a more than 2.5-fold increase in maximum viable cell density. Moreover, supplementation of six different commercial CDMs with 3.6 μM THF yielded up to 2.9-fold enhancement of maximum viable cell density. An anchorage- and serum-dependent DHFR-deficient CHO DG44 cell line was adapted within two consecutive passages to suspension growth in in-house CDM supplemented with 3.6 μM THF. These data indicate that supplementation of chemically defined cell culture media with greater than 0.2 μM THF can help achieve a high density of suspension-adapted DHFR-deficient CHO DG44 cells and may facilitate rapid adaptation of nonadapted DHFR-deficient CHO DG44 cells to suspension culture. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 32:1539-1546, 2016.
authors: Dept. of Chemical Engineering, Hanyang University, Seoul, 133-791, South Korea.
authors: Bong Gyun
authors: BG
authors: Kim
authors: Dept. of Chemical Engineering, Hanyang University, Seoul, 133-791, South Korea.
authors: Hong Woo
authors: HW
authors: Park
mesh_terms: D000818:Animals; D016466:CHO Cells; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D003412:Cricetulus; D003470:Culture Media / Q000737:chemistry / Q000494:pharmacology*; D004305:Dose-Response Relationship, Drug; D013329:Structure-Activity Relationship; D013762:Tetrahydrofolate Dehydrogenase / Q000172:deficiency / Q000378:metabolism*; D013763:Tetrahydrofolates / Q000737:chemistry / Q000494:pharmacology*
chemical_list: D003470:Culture Media; D013763:Tetrahydrofolates; C030371:5,6,7,8-tetrahydrofolic acid; D013762:Tetrahydrofolate Dehydrogenase
keywords: CHO DG44 cells; chemically defined medium; suspension culture; tetrahydrofolate