We offer a broad portfolio of protocols to evaluate natural immune responses specific both in primary cells (human and mouse) and lymphoid cell lines. This is the perfect complement to study in animal models of acute and chronic inflammation, cancer, and other possibilities. Therefore, our in vivo and in vitro models are suitable for preclinical validation of compounds, anti-inflammatory and immunomodulatory drugs in general.
- Available cell types:
– Primary human lymphoid cells and murine cells (lymphocytes T, B, monocytes, granulocytes and NK cells).
– Dendritic cells.
– Primary endothelial cells.
– Transformed Lines: lymphoid, and synovial endothelial.
– Cancer: we have a large panel of tumor cell lines of different origins (Lymphoid, breast, lung, colon, prostate, brain, liver, etc.) for the preclinical study of drugs. These cell lines are commonly used for studies of antitumor compounds and to study their mechanisms of action specific (pharmacological targets).
– Other cell types available (please request information).
- Available tissues
– Human and rat systemic arteries in culture.
– Rat, mouse, chicken and human embryo pulmonary arteries.
– Human ductus.
Listed below are the main tests performed to complement the in vivo studies:
- Inflamatory compounds Testing:
– Determination of prostaglandins and leukotrienes by ELISA; COX-1 and COX- 2 activity.
– Determination of proinflammatory cytokines (ELISA, qRT-PCR and flow cytometry).
– Studies of activation in macrophages (iNOS, COX-2, cytokines, PGs, etc…).
- Cell Biology:
– Metabolism of nitric oxide (NO) and reactive oxygen species (ROS) determination. Analysis of ROS and peroxynitrite
– Proliferation and cell cycle studies in primary cells (lymphocytes).
– Studies of phagocytosis (neutrophils, monocytes and dendritic cells).
– Activity of transcription factors and gene regulation (NF-kB, NF-AT, AP-1, SP1, STAT3).
– Study of the cell cycle (flow cytometry).
– Tests of co-localization and cellular translocation of proteins.
– Determination transcriptional activity of PPARs.
– Tests of invasiveness in Matrigel.
– Room settings and morphometric analysis.
– Uptake of glucose in response to insulin.
– Determination of lipolysis.
– Co-cultured adipocytes / macrophages.
- Cell Signaling:
– MAPKs (ERK 1/2, p38, JKN / SAPK) PKCs; Calcineurin Activity; cyclin activation assays, tyrosine kinases, ubiquitination sumoylation etc.
– Determination of activation markers (flow cytometry).
– Migration assays.
– NK activity.
– Studies of signaling (calcium, pH, identification of molecular targets, etc.).
– Studies on differentiation of 3T3-L1 adipocytes.
- Apoptosis Pathways:
– Potential of mitochondria and release of pro-apoptotic mediators (cytochrome C, AIF).
– Determination of cytotoxicity (MTT, XTT, LDH, propidium iodide), apoptosis and necrosis.
– Study of effector caspase activity.
– DNA Fragmentation (TUNEL).
- Vascular parameters:
– Registration of ionic currents and membrane potential.K and Ca currents in isolated cells (pulmonary and systemic arteries vascular myocytes isolated in fresh and from cultures).
– Registration of intracellular calcium simultaneously with vascular arterial rings (fura-2 fluorescence).
– Registration of vascular Pulmonary and systemic arteries in conductance and resistance.
– Determination of NO and reactive oxygen species. Dihidroetidina (superoxide) and dichlorofluorescein (peroxide), DAF-2 (NO) and other….In arteries, cells or tissue sections.
- Other tests (please request information)