title: Heparin promotes suspension adaptation process of CHO-TS28 cells by eliminating cell aggregation.
abstract: While heparin has been shown to eliminate cell aggregation in suspension adaptations of insect and HEK293 cells for virus-based cell cultures, the role of heparin in long period serum-free suspension adaptation of the anchorage-dependent Chinese hamster ovary (CHO) cell lines remains inconclusive. In this paper, we explore the potential application of heparin in suspension adaptation of CHO cell line which produces an anti-human chimeric antibody cHAb18. Heparin showed a concentration-dependent inhibition of CHO-TS28 cell-to-cell adhesion, with a significant inhibitory effect occurring when the concentration exceeded 250 μg/ml (P < 0.001). Heparin also exhibited a cell aggregation elimination role at all concentrations (P < 0.001). Furthermore, heparin promoted cell growth and antibody secretion, with the highest cell density ((99.83 ± 12.21) × 10(4) cells/ml, P = 0.034) and maximum antibody yield ((9.46 ± 0.94) mg/l, P < 0.001) both occurring at 250 μg/ml heparin. When agitated, cell aggregates were effectively dispersed by 250 μg/ml heparin and a single-cell suspension culture process was promoted. In suspension adapted CHO-TS28 cells, cell growth rates and specific antibody productivity were maintained; while antigen-binding activity improved slightly. Together, our results show that heparin may promote suspension adaptation of anchorage-depended CHO cells by resisting cell aggregation without reducing cell growth, antibody secretion, and antigen-binding activity.
authors.affiliation: Cell Engineering Research Centre & Department of Cell Biology, National Key Discipline of Cell Biology, State Key Laboratory of Cancer Biology, The Fourth Military Medical University, 710032, Xi'an, People's Republic of China. liling25@fmmu.edu.cn
authors.forename: Ling
authors.initials: L
authors.lastname: Li
authors.forename: Jun
authors.initials: J
authors.lastname: Qin
authors.forename: Qiang
authors.initials: Q
authors.lastname: Feng
authors.forename: Hao
authors.initials: H
authors.lastname: Tang
authors.forename: Rong
authors.initials: R
authors.lastname: Liu
authors.forename: Liqing
authors.initials: L
authors.lastname: Xu
authors.forename: Zhinan
authors.initials: Z
authors.lastname: Chen
mesh_terms: D000222:Adaptation, Physiological; D000818:Animals; D016466:CHO Cells; D002448:Cell Adhesion; D002449:Cell Aggregation; D002452:Cell Count; D018929:Cell Culture Techniques / Q000379:methods*; D002460:Cell Line; D006224:Cricetinae; D003412:Cricetulus; D016895:Culture Media, Serum-Free; D006493:Heparin / Q000378:metabolism*
chemical_list: D016895:Culture Media, Serum-Free; D006493:Heparin