Generation and characterisation of an equine macrophage cell line (e-CAS cells) derived from equine bone marrow cells

Werners, Arno; Bull, Sarah; Fink‐Gremmels, Johanna; Bryant, Clare E.

doi:10.1016/j.vetimm.2003.08.012

Cited by 23 publications

(22 citation statements)

References 51 publications

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“…A cell line was chosen rather than primary cells, since it has the advantage of a more uniform response pattern [20]. The well-established human macrophage derived cell line (U937 cells) was used as a reference.…”

Section: Discussionmentioning

confidence: 99%

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Ketamine inhibits LPS-induced tumour necrosis factor-alpha and interleukin-6 in an equine macrophage cell line

et al. 2005

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-Ketamine is widely used in equine anaesthesia. Beside its anaesthetic and analgesic properties, ketamine possesses a cytokine-modulating activity. However, to date, no data are available regarding the inhibitory effect of ketamine on the cytokine response in horses. In horses, cytokines such as tumour necrosis factor-alpha (TNF-α) and interleukin-6 (IL-6) play a pivotal role in the pathogenesis of equine endotoxaemia following gastrointestinal disorders. Hence, the objective of this study was to assess the influence of ketamine on LPS-induced TNF-α and IL-6 formation in an equine macrophage cell line (eCAS cells). The results demonstrate a cytokinemodulating activity of ketamine in an equine cell line, suggesting a beneficial role for ketamine in the treatment of equine endotoxaemia. ketamine / cytokine / equine endotoxaemia / eCAS

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Section: Discussionmentioning

confidence: 99%

“…Two cell lines were used: the equine bone-marrow-derived macrophage cell line (eCAS), developed by Werners et al [20], and the human monoblastoid tumour cell line (U937 cells; ECACC No. 85011440, UK).…”

Section: Cell Lines and Culture Conditionsmentioning

confidence: 99%

Ketamine inhibits LPS-induced tumour necrosis factor-alpha and interleukin-6 in an equine macrophage cell line

et al. 2005

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“…Then, these boxes were placed in a -20°C for 4 h and then in -80°C freezer overnight 16S rRNA using primers (F 5' CGC CTG TTT ATC AAA AAC AT 3' and H 5' CCG GTC TGA ACT CAG ATC ACG T 3') (Ward et al 2005) and COI using primer (F 5' TCA ACC AAC CAC AAA GAC ATT GGC AC 3' and R 5' TAG ACT TCT GGG TGG CCA AAG AAT CA 3') (Palumbi et al 1991). Subsequently, the vials were transferred to a liquid nitrogen can (Werners et al 2004). To recover the cells, the frozen vials were removed from liquid nitrogen and quickly thawed in 37°C water bath, and then the cells were transferred into a flask with complete medium and incubated at 28°C.…”

Section: Cell Cryopreservationmentioning

confidence: 99%

A new epithelial cell line, HBF from caudal fin of endangered yellow catfish, Horabagrus brachysoma (Gunther, 1864)

et al. 2014

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A new epithelial cell line, Horabagrus brachysoma fin (HBF), was established from the caudal fin tissue of yellow catfish, H. brachysoma and characterized. This HBF cell line was maintained in Leibovitz's-15 medium supplemented with 15 % fetal bovine serum (FBS) and subcultured more than 62 times over a period of 20 months. The HBF cell line consists predominantly of epithelial cells and is able to grow at temperatures between 20 and 35°C with an optimum temperature of 28°C. The growth rate of these cells increased as the proportion of FBS increased from 5 to 20 % at 28°C with optimum growth at the concentrations of 15 % FBS. Partial amplification and sequencing of fragments of two mitochondrial genes 16S rRNA and COI confirmed that HBF cell line originated from yellow catfish. The HBF cells showed strong positive reaction to the cytokeratin marker, indicating that it was epithelial in nature. HBF cell line was inoculated with tissue homogenate from juveniles of Sea bass, Lates calcarifer infected with viral nervous necrosis virus (VNNV) and found not susceptible to VNNV. The extracellular products of Vibrio cholerae MTCC 3904 were toxic to the HBF cells. These cells were confirmed for the absence of Mycoplasma sp by PCR.

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“…The equine macrophage cell line developed in the present study was established in the absence of viral transformation, has been in continuous culture for more than three years, and has maintained macrophage-specific properties throughout this time, namely NSE and phagocytosis activity, nitrite oxide production, and the expression of two available equine macrophage-specific markers (CZ2.2, and Ki-M6). Spontaneous immortalization of monocyte derived cell lines has been previously reported with ovine monocytes (Olivier et al, 2001) and in the generation of an equine macrophage (e-CAS) cell line (Werners et al, 2004). Monocytic mammalian cell lines have been established previously by viral infection of or by introduction of viral oncogenes into primary cells (SV40, V-myc and V-raf) (Blasi et al, 1989; Kreuzburg-Duffy and MacDonald, 1994; Nagata et al, 1983).…”

Section: Discussionmentioning

confidence: 90%

Characterization of an equine macrophage cell line: Application to studies of EIAV infection

Fidalgo-Carvalho¹,

Craigo

Barnes

et al. 2009

Veterinary Microbiology

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EIAV is a monocyte/macrophage tropic virus. To date, even though EIAV has been under investigation for numerous years, very few details have been elucidated about EIAV/macrophage interactions. This is largely due to the absence of an equine macrophage cell line that would support viral replication. Herein we describe the spontaneous immortalization and generation of a clonal equine macrophage-like (EML) cell line with the functional and immunophenotype characteristics of differentiated equine monocyte derived macrophage(s) (eMDM(s)). These cells possess strong non-specific esterase (NSE) activity, are able to phagocytose fluorescent bioparticles, and produce nitrites in response to LPS. The EML-3C cell line expresses the EIAV receptor for cellular entry (ELR1) and supports replication of the virulent EIAV PV biological clone. Thus, EML-3C cells provide a useful cell line possessing equine macrophage related properties for the growth and study of EIAV infection as well as of other equine macrophage tropic viruses.

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Generation and characterisation of an equine macrophage cell line (e-CAS cells) derived from equine bone marrow cells

Cited by 23 publications

References 51 publications

Ketamine inhibits LPS-induced tumour necrosis factor-alpha and interleukin-6 in an equine macrophage cell line

Ketamine inhibits LPS-induced tumour necrosis factor-alpha and interleukin-6 in an equine macrophage cell line

A new epithelial cell line, HBF from caudal fin of endangered yellow catfish, Horabagrus brachysoma (Gunther, 1864)

Characterization of an equine macrophage cell line: Application to studies of EIAV infection

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