Analysis of high-pressure-induced disruption of human erythrocytes by flow cytometry.
High-pressure-induced hemolysis is suppressed by pretreating human erythrocytes at 49 degrees C, or enhanced by pretreatment with trypsin. So, the response of these pretreated cells to a pressure of 200 MPa was examined using flow cytometry. In the case of intact erythrocytes, a major product was fragmented particles. From 49 degrees C-pretreated cells, vesicles were mainly released. Trypsin-pretreated cells mainly produced open ghosts. Additionally, intact erythrocytes, 49 degrees C-pretreated ones, and trypsin-pretreated ones also released at 200 MPa vesicles of diameter 464 +/- 9, 259 +/- 18, and 574 +/- 16 nm, respectively. These results suggest that mother cells, fragmented particles, vesicles, and open ghosts from 200 MPa-treated erythrocytes are easily monitored by flow cytometry and that the size of released vesicles may also be an important factor in high-pressure-induced hemolysis.
Yamaguchi
Takeo
T
Department of Chemistry, Faculty of Science, Fukuoka University, Jonan-ku, Fukuoka, 814-0180 Japan.
Terada
Shigeyuki
S
D000399:Air Pressure; D048429:Cell Size / Q000502:physiology*; D004910:Erythrocyte Membrane / Q000502:physiology*; D004912:Erythrocytes / Q000502:physiology*; D005434:Flow Cytometry; D006461:Hemolysis / Q000502:physiology; D006801:Humans; D013049:Spectrin / Q000378:metabolism; D014357:Trypsin / Q000494:pharmacology
D013049:Spectrin; D014357:Trypsin