Measurement of Chemotaxis of Peripheral Monocytes by Agarose Plate Method

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  • Agarose plate法による末梢血単球遊走能の測定
  • Agarose plateホウ ニヨル マッショウケツ タンキュウ ユウソウノ

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Chemotaxis of peripheral monocytes was measured by agarose plate method in children and adults. Mononuclear cells (MNC) were collected from heparinized peripheral blood by Ficoll-Hypaque centrifugation and they were suspended in Hanks balanced salt solution at a concentration of 1×108/ml. The center well on agarose plate received 10 μl of the MNC suspension, and the outer and inner wells received chemotactic factor (zymosan-activated human serum) and non-chemotactic control medium, respectively. The agarose plate was incubated at 37°C in a humidified atmosphere containing 5% CO2 in air for 20 hours. Chemotaxis assay was done by measuring the linear distance the cells had moved from the margin of the well toward the chemotactic factor (chemotaxis) or the control medium (random migration).<br>As the appropriate incubation time was 20 hours and MNC concentration was 1×108/ml, the following studies were done under such conditions. Monocyte chemotaxis was not demonstrated when culture filtrate of E. coli was used as a chemotactic factor. There was no significant difference of monocyte chemotaxis and random migration between children and adults: chemotaxis values (mean±S. D.) were 8.5±0.7 (1∼5 years of age), 8.6±1.5 (6∼11 years), 8.9±1.0 (12∼19 years), and 9.9±1.3 (adults).<br>The method is simple and reproducible and can provide further information as to monocyte functions.

Journal

  • Rinsho Ketsueki

    Rinsho Ketsueki 21 (3), 328-335, 1980

    The Japanese Society of Hematology

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