PAN Olsztyn

• Laminair chamber Thermo Scientific MSC-ADVANTAGE (180),
• Laminair chamber Polon’60,
• Three Laminair chamber Thermo Scientific MSC-ADVANTAGE (120),
• Two Incubator CO₂ ThermoForma,
• Incubator CO₂ Sanyo,
• Two Centrifuge MPW 350R,
• Centrifuge MPW 223E,
• Peristaltic pumps VederFlex Scientific,
• Two Inverted microscope (standard) Olympus CKX41,
• Three Incubator with shaker EB TH15(Edmund Bühler),
• Two Water bath with shaker Julabo SW23,
• Ultrasonic pipette-washer SonoreTwo Autoclave Sanyo;

• Laminair chamber Holten 90,
• Centrifuge Hettich Universal 32R,
• Centrifuge Eppendorf miniSpin,
• Gel electrophoresis device BIO-RAD PowerPac,
• UV Ttransilluminator WEALTEC,
• Thermocycler BIO-RAD MJ Mini,
• Whipper TissueRuptor Qiagen;

• Polyacrylamide gel electrophoresis device BIO-RAD PowerPac,
• Semi-dry transfer system TE70PWR Amersham Biosciences,
• Wet transfer system BIO-RAD PowerPac,
• RollMixer Chemland,
• Protein detection system SNAP i.d. Millipore;

• synchronization of ovulation in cows and pigs, superovulation and insemination,
• ultrasound diagnostic (USG) of reproductive organs in cows, pigs and mares,
• laparotomy of cows and pigs (blood vessels cannulation in reproduction system, corpus luteum microdialysis and other structure),
• intravital acquisition ovaries and corpus luteum (laparotomy and calpotomy),
• uterine mucous membrane biopsies in cows;

Enzymatic isolation and cells culture:

• epithelium and stromal cells from the endometrium of cows, pigs, mares and cats,
• steroidogenic cells from the corpus luteum of cows, mares and cats,
• granulosa cells from the ovarian follicle of cows,
• endothelium cells of corpus luteum and mucous membrane blood vessels in cows, pigs, mares and cats,
• epithelial cells of bovine mammary gland cisternae;

• cows, pigs, mares and cats endometrium explants,
• cows, pigs and mares corpus luteum explants,
• bovine mammary gland cisternae explants;

3.1.    Molecular biology methods:

• pro- and anti apoptotic factor (TNFα, IFNγ, IL-1α, Il-6, FasL, BCL2, BAX, Caspase3, Caspase8) expression and their receptors,
• enzymes steroidogenic pathway (3β-HSD, StAR, CYTp450)expression,
• enzymes of arachidonic acid metabolism pathway expression:
  
  • leukotrienes pathway (5-LOX) and their receptors,
  • prostaglandins pathway (PTGS-2, PGES, PGFS, 9KPGR, AKR1B5),
• oksitocin and their receptors expression,
• nerves growth factor expression and its receptors (TrkA, p75) with used Real Time PCR and semiquantitative RT PCR for analysis of mRNA levels and Western Blot method for analysis of protein level,

• protein hormones (LH, PRL) and their receptors (LH/hCG) with iodine marked specimen used (J-125),
• steroidogenic hormones (P4, A4, T, E1, E2, cortisol), prostaglandins and methabolit of PGF2α – PGFM, with tritium marked hormones (H3) and protein hormones iodination (LH, PRL) and cytokine (IL-1β, IL-6, TNF-α),

• protein hormones (LH),
• steroidogenic hormones (P4, T4, E2, cortisol),
• prostaglandins (PGE2, PGF2α),
• oxytocin and endothelin with hormones marked horseradish peroxidase (HRP) or biotin used;

• arachidonic acid pathway enzymes in ovary, uterus and epithelial cells of mammary gland cisternae (PTGS-2, PGES, PGFS, AKR1B5, 5-LO),
• cytokine in ovary and uterus (TNF-α, IL-1α, IL-1β and others),
• enzymes of cholesterol pathway in ovary (P450scc, 3β-HSD, aromatase) and epithelial cells of mammary gland cisternae (11β-HSD, 3β-HSD),
• receptors (TNFR I, TNFR II, IL-1αR, IL-1 βR, LTRI, LTRII, GC-R, LPAR-1, LPAR-2, LPAR-3, LPAR-4, LPAR4, OTR, TrkA, p75),
• neurotransmitters and their markers (NPY, VIP, SOM, SP, GAL, CGRP, PACAP, DβH, VACHT, nNOS);