title: HEK 293 cell suspension culture using fibronectin-adsorbed polymer nanospheres in serum-free medium.
abstract: Previously, we reported on suspension culture of anchorage-dependent animal cells using plain polymer nanospheres in serum-containing medium. For commercial cell culture, it is more advantageous to use serum-free medium than serum-containing medium. To culture anchorage-dependent animal cells using polymer nanospheres in serum-free medium, the nanospheres need to be coated with cell adhesion proteins. In this study, we utilized fibronectin-adsorbed polymer nanospheres for suspension culture of anchorage-dependent animal cells in serum-free medium. Fibronectin was adsorbed onto poly(lactic-co-glycolic acid) nanospheres (433 nm in average diameter) by immersing the nanospheres in fetal bovine serum. The nanospheres were used to culture human embryonic kidney (HEK) 293 cells in serum-free medium in stirred suspension bioreactors. Nanospheres attached between HEK 293 cells and promoted cell aggregate formation compared with culture without nanospheres. Most cells in the aggregates were viable over a 10-day culture period. Importantly, the use of poly(lactic-co-glycolic acid) nanospheres promoted the cell growth significantly, compared with culture without nanospheres (3.8- vs 1.8-fold growth). The nanosphere culture method developed in this study removes the time-consuming and costly process of adaptation of anchorage-dependent animal cells to suspension culture in serum-free medium. This culture method may be useful for the large-scale suspension culture of various types of anchorage-dependent animal cells in serum-free medium.
authors.affiliation: Department of Chemical Engineering, Hanyang University, Seoul 133-791, Korea.
authors.forename: Ju Hee
authors.initials: JH
authors.lastname: Ryu
authors.forename: Sang-Soo
authors.initials: SS
authors.lastname: Kim
authors.forename: Seung-Woo
authors.initials: SW
authors.lastname: Cho
authors.forename: Cha Yong
authors.initials: CY
authors.lastname: Choi
authors.forename: Byung-Soo
authors.initials: BS
authors.lastname: Kim
mesh_terms: D000818:Animals; D019149:Bioreactors; D002417:Cattle; D002449:Cell Aggregation; D018929:Cell Culture Techniques; D002460:Cell Line; D016895:Culture Media, Serum-Free; D005353:Fibronectins / Q000378:metabolism*; D006801:Humans; D019344:Lactic Acid / Q000737:chemistry / Q000378:metabolism; D008422:Materials Testing; D043942:Nanotubes / Q000737:chemistry* / Q000648:ultrastructure; D011100:Polyglycolic Acid / Q000737:chemistry / Q000378:metabolism; D000077182:Polylactic Acid-Polyglycolic Acid Copolymer; D011108:Polymers / Q000737:chemistry / Q000378:metabolism*
chemical_list: D016895:Culture Media, Serum-Free; D005353:Fibronectins; D011108:Polymers; D000077182:Polylactic Acid-Polyglycolic Acid Copolymer; D011100:Polyglycolic Acid; D019344:Lactic Acid