Comparison of in vitro cell cytotoxic assays for tumor necrosis factor

Flick, David A.; Gifford, George E.

doi:10.1016/0022-1759(84)90147-9

Cited by 697 publications

(303 citation statements)

References 5 publications

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“…3). This differential capacity of LPS induction has also been reported depending on the bacterial origin of the LPS [8,13]. This phenomenon may explain the diminished TNFα levels observed in Figure 2b, after LPS-induction.…”

Section: Discussionsupporting

confidence: 78%

“…As described [8], 100 µL of a suspension of 4 × 10 5 L929 fibroblasts (ATCC CCL 1; American Type Culture Collection, Rockville, MD, USA) were seeded in RPMI 10% FBS in 96-well flat bottomed microtitration plates. The culture medium was eliminated and 100 µL of each supernatant in triplicate, obtained from ex-vivo LPS-stimulated WBCS, were added (three samples for each species).…”

Section: Cytotoxic Assay For Human and Feline Tnfαmentioning

confidence: 99%

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An anti-human recombinant tumor necrosis factor alpha (TNF?) monoclonal antibody recognizes an epitope in feline TNF?

et al. 2003

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-It is likely that the murine response to human recombinant TNFα (hrTNFα) may generate antibodies (Ab) to epitopes present in TNFα from other species. Here, we demonstrate that F5 antihrTNFα monoclonal antibody (mAb) recognizes feline TNFα while E8 anti-hrTNFα mAb failed to do so. In order to demonstrate that E8 and F5 mAb recognize different epitopes in the hrTNFα molecule, a constant concentration of E8 and variable concentrations of F5 were incubated with solid phase bound hrTNFα. Binding of E8 and F5 to hrTNFα was determined with anti-µ and γ chain specific Ab. F5 bound equally to hrTNFα in the presence or absence of E8 and the same amount of E8 bound to hrTNFα, in spite of the presence of F5. When using the E8 and F5 mAb for capturing the TNFα from the equine, canine, feline and bovine species, in supernatants of an ex vivo lipopolysaccharide (LPS)-stimulated whole blood cell culture, we only detected the feline TNFα by F5 mAb (p = 0.001). By a cytotoxic assay on L929 fibroblasts, we indeed demonstrated the feline TNFα production after the LPS stimulus. In an inhibition assay, the human and feline cytokines competed for F5, although the inhibition of native human TNFα binding to F5 was significant but only about 20% (p = 0.001). In conclusion, most likely the F5 anti-hrTNFα mAb recognizes an epitope in feline TNFα. Its immunomodulatory potential in the feline model remains to be studied. monoclonal antibodies / tumor necrosis factor alpha / cross-reactivity

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Section: Discussionsupporting

confidence: 78%

Section: Cytotoxic Assay For Human and Feline Tnfαmentioning

confidence: 99%

An anti-human recombinant tumor necrosis factor alpha (TNF?) monoclonal antibody recognizes an epitope in feline TNF?

et al. 2003

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“…Crystal Violet staining HSC-4 cells in response to Asc or Asc6Palm were quantitated by the Crystal Violet staining (Gillies et al 1976), which is based on the inability of staining of dead cells in contrast to ability of staining of adherent viable cells (Flick and Gifford 1984). The medium in a 24-well plate was aspirated, and HSC-4 cells were stained for 10 min with 0.5% Crystal Violet (Wako Pure Chemical Industries, Ltd., Osaka) dissolved in methanol.…”

Section: Hyperthermia Treatment By Cret Systemmentioning

confidence: 99%

Anticancer effects of 6-o-palmitoyl-ascorbate combined with a capacitive-resistive electric transfer hyperthermic apparatus as compared with ascorbate in relation to ascorbyl radical generation

et al. 2011

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The aim of the present study is to determine the anti-proliferative activity of 6-o-palmitoyl-L-ascorbic acid (Asc6Palm) that is a lipophilic derivative of L-ascorbic acid (Asc), on human tongue squamous carcinoma HSC-4 cells by combined use of hyperthermia in comparison to Asc. Asc6Palm or Asc were administered to HSC-4 cells for 1 h, to which hyperthermia at 42°C was applied for initial 15 min. After further 1-72 h incubation at 37°C, cell proliferation was determined with Crystal Violet staining. Ascorbyl radical (AscR) in HSC-4 cell suspension was measured by electron spin resonance (ESR), and cell morphology was observed with scanning electron microscopy (SEM). At 37°C, 4 mM Asc or 0.35 mM Asc6Palm were enough to suppress proliferation of HSC-4 cells. By combined use of hyperthermia at 42°C, cell proliferation was decreased when compared to 37°C. After Asc of 4 mM was incubated with HSC-4 cell suspensions at 37°C or 42°C for 0-180 min, the signal intensity of ascorbyl radical (AscR) by ESR was not different regardless of the presence or absence of cells at 37°C, whereas AscR signal was enlarged in the presence of HSC-4 cells at 42°C. It was suggested that oxidation of Asc occurred rapidly in HSC-4 cells by hyperthermia, and thereby enhanced the anti-proliferative activity. By SEM observation, the surface of HSC-4 cells treated with Asc6Palm revealed distinct morphological changes. Thus, the combined regimen of Asc6Palm and hyperthermia is expected to exert a marked antitumor activity.

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“…14 At the end of incubation, cell cultures were washed with PBS to remove nonadherent dead cells, and adherent viable cells were then fixed with methanol and stained with 1% crystal violet solution at room temperature for 10 min. Plates were thoroughly washed with water, and crystal violet dye was dissolved in 33% acetic acid.…”

Section: Crystal Violet Testmentioning

confidence: 99%

Novel platinum(IV) complexes induce rapid tumor cell death in vitro

Kaludjerović

Miljković

Momčilović³

et al. 2005

Intl Journal of Cancer

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The anticancer activity of platinum complexes has been known since the discovery of classical Pt(II)‐based drug cisplatin. However, Pt(IV) complexes have greater inertness than corresponding Pt(II) complexes, thus allowing the oral administration and reducing the toxicity associated with platinum‐based chemotherapy. Here, we describe the in vitro antitumor activity of some novel Pt(IV)‐based agents against mouse fibrosarcoma L929 cells and human astrocytoma U251 cells. The cytotoxicity of 2 Pt(IV) complexes with bidentate ethylenediamine‐N,N′‐di‐3‐propanoato esters was found to be markedly higher than that of their Pt(II) counterparts and comparable to the antitumor action of cisplatin. In contrast to cisplatin, which caused oxidative stress‐independent apoptotic cell death of tumor cells, these Pt(IV) complexes induced oxygen radical‐mediated tumor cell necrosis. Importantly, the cytotoxic action of novel Pt(IV) complexes was markedly more rapid than that of cisplatin, indicating their potential usefulness in anticancer therapy. © 2005 Wiley‐Liss, Inc.

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Comparison of in vitro cell cytotoxic assays for tumor necrosis factor

Cited by 697 publications

References 5 publications

An anti-human recombinant tumor necrosis factor alpha (TNF?) monoclonal antibody recognizes an epitope in feline TNF?

An anti-human recombinant tumor necrosis factor alpha (TNF?) monoclonal antibody recognizes an epitope in feline TNF?

Anticancer effects of 6-o-palmitoyl-ascorbate combined with a capacitive-resistive electric transfer hyperthermic apparatus as compared with ascorbate in relation to ascorbyl radical generation

Novel platinum(IV) complexes induce rapid tumor cell death in vitro

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