Acta Univ. Agric. Silvic. Mendelianae Brun. 2006, 54(5), 107-116 | DOI: 10.11118/actaun200654050107

Využití metod detekce apoptózy a nekrózy neutrofilních granulocytů krve skotu in vitro

Petr Sláma1, Zbyšek Sládek1, Dušan Ryšánek2
1 Ústav morfologie, fyziologie a genetiky zvířat, Mendelova zemědělská a lesnická univerzita v Brně, Zemědělská 1, 613 00 Brno, Česká republika
2 Výzkumný ústav veterinárního lékařství v Brně, Hudcova 70, 621 32 Brno, Česká republika

Studie byla in vitro analýzou apoptózy a nekrózy bovinních neutrofilů krve, která byla založena na detekci jejich morfologických a biochemických znaků. Experiment byl proveden na šesti klinicky zdravých jalovicích kříženkách holštýnského a českého strakatého plemene ve věku 16-18 měsíců. Čerstvá krev a krev kultivovaná in vitro byly analyzovány světelnou a elektronovou mikroskopií a průtokovou cytometrií. Pomocí světelné mikroskopie nebyly v čerstvé krvi zjištěny apoptotické ani nekrotické neutrofily. Naproti tomu bylo pomocí průtokové cytometrie zaznamenáno 3,34 % apoptotických a 0,32 % nekrotických neutrofilů. Po čtyřech hodinách inkubace bylo v krvi pomocí světelného mikroskopu zjištěno 3,53 % apoptotických a 1,37 % nekrotických neutrofilů. Za stejných podmínek vykazovaly vzorky krve v průtokovém cytometru 17,54 % apoptotických a 0,58 % nekrotických neutrofilů. Korelační koeficient v podílu apoptotických neutrofilů mezi světelnou mikroskopií a průtokovou cytometrií byl 0,625 (P

apoptóza, nekróza, neutrofil, fosfatidylserin, průtoková cytometrie

Application of methods for detection of apoptosis and necrosis of bovine blood neutrophil granulocytes in vitro

The study was an in vitro analysis of bovine blood neutrophil apoptosis and necrosis based on the detection of their morphological and biochemical features. The experiment was carried out in six clinically healthy Holstein x Bohemian Red Pied crossbred virgin heifers aged 16 to 18 months. The fresh and in vitro cultivated blood were analysed by light and electron microscopy and flow cytometry. In fresh blood, non apoptotic or necrotic neutrophils were found by light microscopy. On the other hand, 3.34% apoptotic and 0.32% necrotic neutrophils were detected by flow cytometry. After four hours of incubation, 3.53% apoptotic and 1.37% necrotic neutrophils were found by light microscopy. In the same conditions, there were assessed 17.54% apoptotic and 0.58% necrotic neutrophils by flow cytometry. Correlation coefficient between light microscopy and flow cytometry in portion of apoptotic neutrophils was 0.625 (P

Keywords: apoptosis, necrosis, neutrophil, phosphatidylserine, flow cytometry
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Tato práce vznikla za podpory výzkumného projektu GA ČR č. 524/03/1531.

Received: July 20, 2006; Published: December 3, 2014  Show citation

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Sláma, P., Sládek, Z., & Ryšánek, D. (2006). Application of methods for detection of apoptosis and necrosis of bovine blood neutrophil granulocytes in vitro. Acta Universitatis Agriculturae et Silviculturae Mendelianae Brunensis, 54(5), 107-116. doi: 10.11118/actaun200654050107
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