The DF-1 Chicken Fibroblast Cell Line: Transformation Induced by Diverse Oncogenes and Cell Death Resulting from Infection by Avian Leukosis Viruses

Himly, Martin; Foster, Douglas N.; Bottoli, Ivan; Iacovoni, Jason S.; Vogt, Peter K.

doi:10.1006/viro.1998.9290

Cited by 381 publications

(294 citation statements)

References 64 publications

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“…Mammalian cells are refractory to the action of most single oncogenes; in contrast, avian cells are highly sensitive indicators of the oncogenic potential of single agents. They can be readily transformed by single oncoproteins, for example, Myc, Jun, Qin, P3K or Akt, all of which are either inactive or extremely inefficient as single oncogenic agents in mammalian cell systems (Bister et al, 1977;Chang et al, 1997;Aoki et al, 1998;Himly et al, 1998).…”

Section: Oncogenic Transformations By Rheb Mutants H Jiang and Pk Vogtmentioning

confidence: 99%

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Constitutively active Rheb induces oncogenic transformation

Jiang

Vogt

2008

Oncogene

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Section: Oncogenic Transformations By Rheb Mutants H Jiang and Pk Vogtmentioning

confidence: 99%

“…Infected cells were fed every other day with nutrient agar for 14 days and stained with crystal violet. The ability of cells to grow in soft nutrient agar suspension was tested according to published techniques (Himly et al, 1998;Cohen et al, 2001;Sonderegger and Vogt, 2003). Colonies were counted after 14 days of incubation.…”

Section: Plasmid Constructionmentioning

confidence: 99%

Constitutively active Rheb induces oncogenic transformation

Jiang

Vogt

2008

Oncogene

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“…15,23 Briefly, virus propagation was initiated by transfection of retroviral plasmid constructs into the DF-1 chick fibroblast cell line. 16,17 After several passages of DF-1 cells, the supernatants were collected and concentrated by centrifugation at 22 000 rpm for 3 h in a Beckman SW 28 rotor and stored in aliquots at À801C. Typical virus titers were 1-5 Â 10 8 cfu/ml.…”

Section: Constructs and Virus Generationmentioning

confidence: 99%

“…16,17 A few DF-1 cells appeared to undergo PCD 24 h after the conventional transfection procedure of viral constructs expressing human Bax (hBax). Following several passages, however, DF-1 cells expressing hBax or its fusion protein with enhanced green fluorescent protein (EGFP) could be cultured normally in growth medium although hBax or EGFP-Bax were abundantly expressed in DF-1 cells (Figure 1b).…”

Section: Introductionmentioning

confidence: 99%

Distinct susceptibility of developing neurons to death following Bax overexpression in the chicken embryo

et al. 2005

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Bax is a proapoptotic protein that is required for programmed cell death (PCD) of many neuronal populations. Here we show that, during an early period of retinal PCD and in naturally occurring sensory and motor neuron (MN) death in the spinal cord, Bax delivery results in enhanced death of these neural populations. In contrast, Bax overexpression fails to enhance an early phase of MN death that occurs in the cervical spinal cord, although overexpressed Bax appears to be activated in dying MNs. Bax overexpression does not also affect the survival of immature neurons prior to the PCD period. Taken together, these data provide the first in vivo evidence suggesting that Bax appears to act selectively as an executioner only in neurons undergoing PCD. Furthermore, although Bax appears to mediate the execution pathway for PCD, the effect of Bax overexpression on susceptibility to death differs between different neuronal populations.

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“…DF-1 cells are an immortalized avian cell line spontaneously derived from ev-0 fibroblasts of 10-day-old chicken embryos (13). ev-0 cells do not contain endogenous ALV sequences, and thus these cells can be used as the substrates for the detection of exogenous ALV and other avian retroviruses (2,14).…”

mentioning

confidence: 99%

Detection of Avian Retroviruses in Vaccines by Amplification on DF-1 Cells with Immunostaining and Fluorescent Product-Enhanced Reverse Transcriptase Endpoint Methods

et al. 2013

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In order to ensure the safety of vaccines produced on avian cells, rigorous testing for the absence of avian retroviruses must be performed. Current methods used to detect avian retroviruses often exhibit a high invalid-test/false-positive rate, rely on hardto-secure reagents, and/or have readouts that are difficult to standardize. Herein, we describe the development and validation of two consistent and sensitive methods for the detection of avian retroviruses in vaccines: viral amplification on DF-1 cells followed by immunostaining for the detection of avian leukosis virus (ALV) and viral amplification on DF-1 cells followed by fluorescent product-enhanced reverse transcriptase (F-PERT) for the detection of all avian retroviruses. Both assays share an infectivity stage on DF-1 cells followed by a different endpoint readout depending on the retrovirus to be detected. Validation studies demonstrated a limit of detection of one 50% cell culture infectious dose (CCID 50 )/ml for retrovirus in a 30-ml test inoculum volume for both methods, which was as sensitive as a classical method used in the vaccine industry, namely, viral amplification on primary chicken embryo fibroblasts followed by the complement fixation test for avian leukosis virus (COFAL). Furthermore, viral amplification on DF-1 cells followed by either immunostaining or F-PERT demonstrated a sensitivity that exceeds the regulatory requirements for detection of ALV strains. A head-to-head comparison of the two endpoint methods showed that viral amplification on DF-1 cells followed by F-PERT is a suitable method to be used as a stand-alone test to ensure that vaccine preparations are free from infectious avian retroviruses.V accines are very effective at reducing death and suffering caused by viral diseases. Many vaccines are generated on primary avian cells, such as the vaccines for yellow fever, measles, and mumps. International health authorities require vaccines produced on avian cells to be tested for and demonstrated to be free of infectious avian retroviruses such as avian leukosis virus (ALV) and reticuloendotheliosis virus (REV) (1, 2, 3, 4). For example, detailed regulatory requirements are outlined specifically for the detection of ALV and REV in the European Pharmacopeia (Ph. Eur.) (2). Furthermore, the detection of reverse transcriptase (RT) activity, which is indicative of the presence of retroviruses, by highly sensitive PCR-based methods is recommended by several international regulatory authorities (3, 4, 5).The detection of RT activity in yellow fever, measles, and mumps vaccines (6, 7) clearly demonstrated a need for effective methods to test for the presence of replication-competent retroviral contaminants. It was discovered upon further investigation that nonreplicative endogenous avian retroviral elements not associated with infectious retroviruses were responsible for the positive RT signals observed in these vaccines (8,9,10,11,12). As a consequence, it is now understood that it is necessary to test for the presence of infectious ...

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The DF-1 Chicken Fibroblast Cell Line: Transformation Induced by Diverse Oncogenes and Cell Death Resulting from Infection by Avian Leukosis Viruses

Cited by 381 publications

References 64 publications

Constitutively active Rheb induces oncogenic transformation

Constitutively active Rheb induces oncogenic transformation

Distinct susceptibility of developing neurons to death following Bax overexpression in the chicken embryo

Detection of Avian Retroviruses in Vaccines by Amplification on DF-1 Cells with Immunostaining and Fluorescent Product-Enhanced Reverse Transcriptase Endpoint Methods

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