title: Bioactive recombinant human lactoferrin, derived from rice, stimulates mammalian cell growth.
abstract: Today there is a concern about the use of animal source proteins and peptides in cell culture applications due to potential contamination by adventitious infectious pathogens. Recombinant production of these proteins using a plant host provides a safe and cost effective alternative. In this paper, we tested the effect of rice-derived recombinant human lactoferrin (rhLF) on mammalian cell growth. The purified rhLF was partially (about 50%) iron-saturated (pis-rhLF). Chemical modification of pis-rhLF generated apo-rhLF (<10% iron saturation) or holo-rhLF (>90% iron saturation). All three forms of rhLF (pis, apo, holo) promoted growth of intestinal cells (HT-29) measured as [(3)H]-thymidine incorporation or viable cell count, but holo-rhLF was most effective. Holo-rhLF was further tested on hybridoma, osteoblast, and human embryonic kidney cells. Results showed that holo-rhLF promoted cell growth and reduced cell doubling time. The concentration of holo-rhLF in media was critical in promoting cell growth and each cell line had different concentration dependence with the most effective range from 5 to 200 mg/L. The effect of rhLF on antibody production was determined using a hybridoma cell line. Significantly, more antibodies were produced by cells grown with holo-rhLF than cells grown without holo-rhLF. We also compared the effect of holo-rhLF to that of human transferrin, a component commonly used in cell culture media as an iron source. Holo-rhLF was as effective as human transferrin in promoting cell growth and antibody production. Considering all the data obtained, we conclude that rhLF from rice is effective in promoting mammalian cell growth and increasing cell productivity.
authors.affiliation: InVitria, Davis, CA 95618, USA. NHuang@invitria.com
authors.forename: N
authors.initials: N
authors.lastname: Huang
authors.forename: D
authors.initials: D
authors.lastname: Bethell
authors.forename: C
authors.initials: C
authors.lastname: Card
authors.forename: J
authors.initials: J
authors.lastname: Cornish
authors.forename: T
authors.initials: T
authors.lastname: Marchbank
authors.forename: D
authors.initials: D
authors.lastname: Wyatt
authors.forename: K
authors.initials: K
authors.lastname: Mabery
authors.forename: R
authors.initials: R
authors.lastname: Playford
mesh_terms: D000042:Absorption / Q000187:drug effects; D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis; D002452:Cell Count; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D005433:Flour; D019073:HT29 Cells; D006801:Humans; D006825:Hybridomas / Q000166:cytology / Q000187:drug effects; D007501:Iron / Q000378:metabolism; D007781:Lactoferrin / Q000494:pharmacology*; D008322:Mammals / Q000378:metabolism*; D012275:Oryza / Q000737:chemistry*; D010006:Osteoblasts / Q000166:cytology / Q000187:drug effects; D051381:Rats; D011994:Recombinant Proteins / Q000494:pharmacology*; D013936:Thymidine / Q000378:metabolism
chemical_list: D000911:Antibodies, Monoclonal; D011994:Recombinant Proteins; D007501:Iron; D007781:Lactoferrin; D013936:Thymidine