CELLULAR MODELS
In vitro models are performed to assess the anti-inflammatory and anti-allergic activity of different compounds.
Cell lines, as well as canine mature cells, provide a useful tool to perform morphofunctional studies on allergic and inflammatory processes.
CELLULAR MODELS
Primary cultures
Isolation and culture of canine skin cells (fibroblasts, mast cells, keratinocytes...) We perform studies on the secretory response of mast cells to different immunologic and non-immunologic stimuli, as well as on the ability of different drugs to inhibit or induce an allergic reaction.
Isolation and culture of canine endothelial cells of the dog's carotid artery allow the study of the expression of adhesion molecules in their membrane, and the evaluation of the efficacy of anti- inflammatory drugs.
Cell lines
Supplied from different research groups, Univet disposes of several canine and human cell lines to perform morphofunctional studies on allergic and inflammatory processes.
| ORIGIN |
CELL TYPE |
SPECIE |
| Mastocytoma |
C2 cell line |
Canine |
| Mastocytoma |
BR cell line |
Canine |
| Mastocytoma |
HMC-1 cell line |
Human |
| Mastocytoma |
MPT-1 cells |
Canine |
SKIN EQUIVALENT MODEL
Complex cellular model, which incorporates the basic cell components of the dog
skin, fibroblasts and keratinocytes, useful tool to investigate the biology and
pathology of canine skin and also to replace animal testing.
Skin equivalents developed from canine cutaneous cells presented a similar
morphological structure than healthy canine skin.
This model has been developed by the research department of UNIVET.
INFLAMMATORY MEDIATORS
IgE mediated mast cell activation is the earliest phenomenon in
an allergic reaction. The mediators released by mast cells can
be assessed by different studies. UNIVET performs:
Determination of inflammatory mediators released by canine
mast cells when stimulated by immunologic and nonimmunologic
agents.
Isolation and determination of
inflammatory molecules expression at
protein level as well as DNA and RNA
levels.
Analysis of adhesion molecules after
the interaction between canine mast
cells and endothelial cells in allergic
processes (Torres et al, 2002):
| MEDIATORS ASSESSED |
IDENTIFICATION / ASSESSMENT |
| Histamine |
ELISA |
| Prostaglandins and leukotriens |
Immunoblot |
| Cytokins (IL-1, TNF- ) |
PCR |
| Adhesion molecules |
Immunocytochemistry
(confocal microscopy, electronic microscopy) |
| |
Immunohystochemistry |
| |
FACS |
OXIDATIVE STRESS MODELS
Inflammatory cells also release reactive oxygen species (ROS)
into their close environment with potentially adverse effects on
tissues. Those ROS released by neutrophils are involved in cell
apoptosis, gene expression regulation, activation of metalloproteases
and cartilage breakdown.
Oxidative stress can be generated in dog cells in vitro and also
ex vivo. This model allows to study the protective antioxidant
effect of different pharmaceutical or dietary compounds to reduce
the oxidative damage at DNA level, to prevent cellular apoptosis
and to reduce cytotoxicity caused by several diseases, drugs or
environmental toxicants:
| DNA fragmentation |
Cytotoxicity |
Apoptosis |
| (COMET assay) |
(MTT assay) |
(FACS, annexin V...) |
FUNCTIONAL ANTI-OXIDATIVE MODELS
Study of the antioxidant effect of different compounds can be
assessed in canine mononuclear cells isolated from blood, after
oral treatment with antioxidant agents:
- Cytotoxicity, apoptosis and DNA damage
- Lymphocyte proliferation