title: Establishment and characterization of duck embryo epithelial (DEE) cell line and its use as a new approach toward DHAV-1 propagation and vaccine development.
abstract: The primary cell culture was derived from duck embryonic tissue, digested with collagenase type I. The existence of cell colonies with epithelial-like morphology, named duck embryo epithelial (DEE), were purified and optimally maintained at 37°C in M199 medium supplemented with 5% fetal bovine serum. The purified cells were identified as epithelial cell line by detecting Keratin-18 expression using immunofluorescence assay. Our findings demonstrated that DEE cell line can be propagated in culture with (i) a great capacity to adhere, (ii) a great proliferation activity, and (iii) a population doubling time of approximately 18h. Chromosomal features of the DEE cell line were remained constant after the 50th passage. Further characterizations of DEE cell line showed that cell line can normally be grown even after several passages and never converted to tumorigenic cells either in vitro or in vivo study. Susceptibility of DEE cell line was determined for transfection and duck hepatitis A type 1 virus (DHAV-1)-infection. Interestingly, the 50% egg lethal dose (ELD50) of the propagated virus in DEE cell line was higher than ELD50 of the propagated virus in embryonated eggs. Finally, DEE cell line was evaluated to be used as a candidate for DHAV-1 vaccine development. Our results showed that the propagated DHAV-1 vaccine strain SDE in DEE cell line was able to protect ducklings against DHAV-1 challenge. Taken together, our findings suggest that the DEE cell line can serve as a valuable tool for DHAV-1 propagation and vaccine production.
authors.affiliation: Shandong Binzhou Animal Science & Veterinary Medicine Academy, 256600 Binzhou, Shandong, China; Shandong Lvdu Bio-Sciences &Technology Co. Ltd., 256600 Binzhou, Shandong, China. Electronic address: wwx1997@126.com.
authors.forename: Wenxiu
authors.initials: W
authors.lastname: Wang
authors.affiliation: Parasitology and Animal Diseases Department, National Research Center, Dokki, Giza, Egypt; Vaccine and Infectious Disease Organization (VIDO), 120 Veterinary Road, University of Saskatchewan, S7N 5E3 Saskatoon, Saskatchewan, Canada. Electronic address: abdo.said@usask.ca.
authors.forename: Abdelrahman
authors.initials: A
authors.lastname: Said
authors.affiliation: Shandong Binzhou Animal Science & Veterinary Medicine Academy, 256600 Binzhou, Shandong, China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Shandong Binzhou Animal Science & Veterinary Medicine Academy, 256600 Binzhou, Shandong, China.
authors.forename: Qiang
authors.initials: Q
authors.lastname: Fu
authors.affiliation: Shandong Binzhou Animal Science & Veterinary Medicine Academy, 256600 Binzhou, Shandong, China.
authors.forename: Yueqiang
authors.initials: Y
authors.lastname: Xiao
authors.affiliation: Shandong Binzhou Animal Science & Veterinary Medicine Academy, 256600 Binzhou, Shandong, China.
authors.forename: Sufang
authors.initials: S
authors.lastname: Lv
authors.affiliation: Shandong Binzhou Animal Science & Veterinary Medicine Academy, 256600 Binzhou, Shandong, China; Shandong Lvdu Bio-Sciences &Technology Co. Ltd., 256600 Binzhou, Shandong, China. Electronic address: bzshenzq@163.com.
authors.forename: Zhiqiang
authors.initials: Z
authors.lastname: Shen
mesh_terms: D000818:Animals; D002448:Cell Adhesion; D002460:Cell Line; D049109:Cell Proliferation; D003470:Culture Media / Q000737:chemistry; D004372:Ducks; D004625:Embryo, Nonmammalian; D004847:Epithelial Cells / Q000502:physiology / Q000821:virology; D042822:Genomic Instability; D004371:Hepatitis Virus, Duck / Q000254:growth & development* / Q000472:pathogenicity; D006524:Hepatitis, Viral, Animal / Q000517:prevention & control; D014761:Viral Hepatitis Vaccines / Q000276:immunology / Q000302:isolation & purification*; D014776:Virus Cultivation / Q000379:methods*
chemical_list: D003470:Culture Media; D014761:Viral Hepatitis Vaccines
keywords: DHAV-1 propagation; Duck embryo epithelial (DEE) cell line; Vaccine development