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Titolo:
Separation of cardiomyocytes and coronary endothelial cells for cell-specific RT-PCR
Autore:
Preisig-Muller, R; Schnitzler, MMY; Derst, C; Daut, J;
Indirizzi:
Univ Marburg, Inst Normale & Pathol Physiol, D-35037 Marburg, Germany UnivMarburg Marburg Germany D-35037 l Physiol, D-35037 Marburg, Germany
Titolo Testata:
AMERICAN JOURNAL OF PHYSIOLOGY-HEART AND CIRCULATORY PHYSIOLOGY
fascicolo: 1, volume: 46, anno: 1999,
pagine: H413 - H416
SICI:
0363-6135(199907)46:1<H413:SOCACE>2.0.ZU;2-8
Fonte:
ISI
Lingua:
ENG
Soggetto:
HEART;
Keywords:
endothelin; troponin T; capillary endothelium; cell picker;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
8
Recensione:
Indirizzi per estratti:
Indirizzo: Preisig-Muller, R Univ Marburg, Inst Normale & Pathol Physiol, Deutschhausstr 2, D-35037 Marburg, Germany Univ Marburg Deutschhausstr 2 Marburg Germany D-35037 y
Citazione:
R. Preisig-Muller et al., "Separation of cardiomyocytes and coronary endothelial cells for cell-specific RT-PCR", AM J P-HEAR, 46(1), 1999, pp. H413-H416

Abstract

A simple method for analyzing the differential gene expression of coronaryendothelial cells and cardiac muscle cells was developed. Cells were isolated from guinea pig hearts by collagenase digestion. In the diluted cell suspension, single cardiomyocytes and capillary fragments containing 6-15 endothelial cells could be identified morphologically. A simple "cell picker" was constructed using a polyethylene pipette with a tip diameter of similarto 150 mu m that was attached to a micromanipulator and connected to an electric miniature valve. Intermittent suction pulses (1- to 2-cm water column) were applied by opening the valve for 100-200 ms at 1-s intervals. Cardiomyocytes (800-1,000) or capillary fragments (150) were picked under visualcontrol using an inverted microscope. The cells were transferred to a reaction tube for RNA extraction, reverse transcription (RT), and DNA amplification (RT-PCR) with gene-specific and intron-spanning primers. All PCR products were verified by sequencing. Troponin T and endothelin-l were found to be specific markers for guinea-pig cardiac muscle cells and coronary endothelial cells, respectively.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 02/12/20 alle ore 18:36:22