Apoptotic effects of hydrogen peroxide and vitamin C on chicken embryonic fibroblasts: redox state and programmed cell death

Abstract

The pro-apoptotic effects of hydrogen peroxide and the purported anti-apoptotic effects of Vitamin C on chicken embryonic fibroblasts were investigated. Hydrogen peroxide induced morphological changes in a dose dependent manner, and a myriad of autophagosomes were observed using transmission electron microscopy. Doxorubicin elicited alterations were not inhibited by co-incubation with Vitamin C except that mitochondrial structure was slightly improved. TUNEL assay, cytotoxicity analysis and flow cytometry revealed that the cytotoxicity, DNA fragmentation and apoptotic rates were dose dependent upon treatment with hydrogen peroxide. Calcium homeostasis was disrupted in a dose dependent manner, and cell cycle was blocked at G(2)/M checkpoint at low concentration and S/G(2) checkpoint at high concentration respectively upon treatment with hydrogen peroxide. The administration of Vitamin C only has a modest effect against doxorubicin induced apoptosis, calcium homeostasis disruption and cell cycle arrest. This research demonstrated that the elevation of reactive oxygen species is sufficient to induce the apoptosis of chicken embryonic fibroblasts, whereas the administration of Vitamin C does not necessarily have certain anti-apoptotic effects, especially when the stimulus is not directly linked with redox state.

Enhanced replication of avian-origin H3N2 canine influenza virus in eggs, cell cultures and mice by a two-amino acid insertion in neuraminidase stalk

“…A/canine/Jiangsu/06/2011(H3N2) identified from pet dogs in the Jiangsu province of China [ 12 ] was used as the wild-type canine influenza virus in this study. Primary cultures of chicken embryo fibroblasts (CEF) were prepared from specific-pathogen-free (SPF) chicken embryos (10 days old) as described previously [ 18 ]. Primary canine bronchiolar epithelial cells (CBE) was prepared from beagles according to protocols previously described [ 19 ].…”

Integration of ALV into CTDSPL and CTDSPL2 genes in B-cell lymphomas promotes cell immortalization, migration and survival

“…Cells were seeded at 0.8 × 10 6 cells in a 10 cm dish at time 0. To induce apoptosis, cells were treated with 50 μM H 2 O 2 as described [ 40 ]. After 48 hours, cells were collected and counted using a BioRad automated cell counter (BioRad TC20) to determine change in cell survival relative to CEFs infected with empty viral vector.…”

Selection of reference genes for quantitative real-time PCR analysis in chicken embryo fibroblasts infected with avian leukosis virus subgroup J

“…Primary cultures of chicken embryo fibroblasts (CEF) were prepared from 10-day-old specific-pathogen-free (SPF) chicken embryos (Yebio Bioengineering Co. Ltd, Qindao, China) as described previously [ 12 ], and maintained in DMEM supplemented with 10% fetal bovine serum (FBS). The cells were seeded (approximately 5 × 10 6 cells/well) in 24-well culture plates.…”