Microarray analysis of early and late passage chicken embryo fibroblast cells

Abstract

Primary cultured cells derived from normal tissue have a limited lifespan due to replicative senescence and show distinct phenotypes such as irreversible cell cycle arrest and enlarged morphology. Studying senescence-associated genetic alterations in chicken cells will provide valuable knowledge of cellular growth characteristics, when compared with normal and rapidly growing cell lines. Microarray analysis of early- and late-passage (passage 4 and 18, respectively) primary chicken embryo fibroblast (CEF) cells was performed with a 4X44K chicken oligo microarray. A total of 1,888 differentially expressed genes were identified with a 2-fold level cutoff that included 272 upregulated and 1,616 downregulated genes in late-passage senescent CEF cells. Bioinformatic analyses were performed using Ingenuity Pathway Analysis (IPA, http://www.ingenuity.com). Of the 1,888 differentially expressed genes in senescent CEF cells, 458 were identified as functionally known genes and only 61 genes showed upregulation. Because senescent cells generally showed the deactivated states of most cellular mechanisms for proliferation and energy metabolism, intensified analysis on upregulated genes revealed that the molecular mechanisms in senescent CEF cells are characterized by the suppression of cell cycle and proliferation, progression of cell death including apoptosis, and increased expression of various secreting factors. These regulatory pathways may be opposite to those found in the immortal CEF cell line, such as the DF-1 immortal line. Further comparison of differentially expressed genes between senescent and immortal DF-1 CEF cells showed that 35 genes overlapped and were oppositely regulated. The global gene expression profiles may provide insight into the cellular mechanisms that regulate cellular senescence and immortalization of CEF cells.

Animal Cell Lines as Expression Platforms in Viral Vaccine Production: A Post Covid-19 Perspective

“…CEF cultures have been involved in vaccine production for many years because they can be easily obtained from SPF eggs. , M199, which is one of the general basal media developed for cell cultures as it has developed parallel to the evolution of cell culture technology, was actually developed for the use of CEF cells in serum-free vaccine production . CEF culture for vaccine production is created by taking the embryo from 10 to 12 day old SPF eggs with aseptic techniques, first dissociate it with mechanical and then enzymatic processes, and then placing the resulting cells in culture dishes and culture them. ,, Compared to egg-based production, the production of more doses of vaccine is achieved by sacrificing a single embryo. It has been observed that the CEF culture, which must be re-established at the end of a certain production cycle, can be passaged 20–22 times before senescence .…”

“…CEF culture for vaccine production is created by taking the embryo from 10 to 12 day old SPF eggs with aseptic techniques, first dissociate it with mechanical and then enzymatic processes, and then placing the resulting cells in culture dishes and culture them. ,, Compared to egg-based production, the production of more doses of vaccine is achieved by sacrificing a single embryo. It has been observed that the CEF culture, which must be re-established at the end of a certain production cycle, can be passaged 20–22 times before senescence . However, in the guidelines published by organisations that determine the regulations in vaccine production such as FDA, EMA, ICH, and World Health Organization (WHO), it is not recommended to passage CEF cultures to be used in vaccine production more than 3 times. ,, The reason for this is the decrease in cellular proliferation as the cell character changes after the number of passages and the resulting decrease in virus production efficiency. , …”

“…It has been observed that the CEF culture, which must be re-established at the end of a certain production cycle, can be passaged 20–22 times before senescence . However, in the guidelines published by organisations that determine the regulations in vaccine production such as FDA, EMA, ICH, and World Health Organization (WHO), it is not recommended to passage CEF cultures to be used in vaccine production more than 3 times. ,, The reason for this is the decrease in cellular proliferation as the cell character changes after the number of passages and the resulting decrease in virus production efficiency. , …”

“… 113 CEF culture for vaccine production is created by taking the embryo from 10 to 12 day old SPF eggs with aseptic techniques, first dissociate it with mechanical and then enzymatic processes, and then placing the resulting cells in culture dishes and culture them. 5 , 11 , 112 Compared to egg-based production, the production of more doses of vaccine is achieved by sacrificing a single embryo. It has been observed that the CEF culture, which must be re-established at the end of a certain production cycle, can be passaged 20–22 times before senescence.…”

Antiproliferative effect and characterization of a novel antifungal peptide derived from human Chromogranin A

“…A549 lung cancer cells (American Type Culture Collection, Manassas, VA, USA) were maintained at the Henan University of Technology (Zhengzhou, China) in RPMI-1640 medium (Gibco-BRL, Grand Island, NY, USA) supplemented with 10% fetal bovine serum (Hyclone, Rockford, IL, USA), with 100 U/ml penicillin and 100 µg/ml streptomycin at 37˚C in a humidified atmosphere containing 5% CO 2 . The normal primary chicken embryo fibroblasts (CEFs) from specific pathogen-free chickens (Experimental Animal Center, Jenan University of Technology, Zhengzhou, China) were isolated using a standard protocol described elsewhere (26). Fibroblasts were grown as a monolayer in Dulbecco's modified Eagle's medium (Gibco-BRL) containing 10% fetal bovine serum (Hyclone).…”