M F Forman1, G L Brower, J S Janicki. 1. Department of Anatomy, Physiology and Pharmacology, Auburn University, Auburn, AL 36849-5517, USA.
Abstract
OBJECTIVE: To test the hypothesis that spontaneous release of histamine occurring during an isolation protocol may modify responses of rat cardiac mast cells (connective tissue-type mast cells) to secretagogues. METHODS: We assessed two protocols for enzymatic dispersion utilizing collagenase, hyaluronidase, and deoxyribonuclease; with protease (Protocol 1, n = 8) or without protease (Protocol 2, n = 3). Spontaneous release of histamine was quantified following mechanical and enzymatic dispersion of the whole heart. RESULTS: Total histamine loss (Mean +/- SEM) was 963+/-92 and 833+/-60 ng/g of tissue weight following Protocols 1 and 2. Percentages of histamine release from cell isolates following Protocol 1 were 40+/-5%, 41+/-6%, and 51+/-7% at 0, 30, and 300 microg/mL of compound 48/80. CONCLUSIONS: Enzymatic dispersion of cardiac mast cells affects their response to secretagogues.
OBJECTIVE: To test the hypothesis that spontaneous release of histamine occurring during an isolation protocol may modify responses of rat cardiac mast cells (connective tissue-type mast cells) to secretagogues. METHODS: We assessed two protocols for enzymatic dispersion utilizing collagenase, hyaluronidase, and deoxyribonuclease; with protease (Protocol 1, n = 8) or without protease (Protocol 2, n = 3). Spontaneous release of histamine was quantified following mechanical and enzymatic dispersion of the whole heart. RESULTS: Total histamine loss (Mean +/- SEM) was 963+/-92 and 833+/-60 ng/g of tissue weight following Protocols 1 and 2. Percentages of histamine release from cell isolates following Protocol 1 were 40+/-5%, 41+/-6%, and 51+/-7% at 0, 30, and 300 microg/mL of compound 48/80. CONCLUSIONS: Enzymatic dispersion of cardiac mast cells affects their response to secretagogues.
Authors: Scott P Levick; Giselle C Meléndez; Eric Plante; Jennifer L McLarty; Gregory L Brower; Joseph S Janicki Journal: Cardiovasc Res Date: 2010-08-24 Impact factor: 10.787
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