RAPID QUANTITATION OF NEUTROPHIL CHEMOTAXIS - USE OF A POLYVINYLPYRROLIDONE-FREE POLYCARBONATE MEMBRANE IN A MULTIWELL ASSEMBLY
RAPID QUANTITATION OF NEUTROPHIL CHEMOTAXIS - USE OF A POLYVINYLPYRROLIDONE-FREE POLYCARBONATE MEMBRANE IN A MULTIWELL ASSEMBLY
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DOI：
10.1016/0022-1759(80)90179-9
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发表时间：
1980-01-01
影响因子：
2.2
通讯作者：
LEONARD, EJ
中科院分区：
文献类型：
--
作者：
HARVATH, L;FALK, W;LEONARD, EJ
开发了一种[人]嗜中性粒细胞趋化性试验，可快速定量评估膜滤器的迁移。测定中的关键因素是使用10 μ m厚的聚碳酸酯膜而没有通常的聚乙烯吡咯烷酮涂层。迁移的中性粒细胞保持粘附在未涂覆的膜上，而20-50%从聚乙烯吡咯烷酮涂覆的膜上脱落。该方法的一个主要优点是，中性粒细胞趋化性可以很容易地量化，因为迁移的细胞粘附在膜表面，并在1个光学平面计数。A 25 mm ×在48孔微趋化性组件中使用80 mm膜片，其每个测定孔仅需要20，000个嗜中性粒细胞和25 μ l引诱剂。在37 ℃下10-20分钟内完成中性粒细胞趋化。C，其中20-30%的细胞迁移到N-甲酰基-甲硫氨酰基-亮氨酰基-苯丙氨酸，40-50%迁移到C5 a [补体成分5的片段]。
A [human] meutrophil chemotaxis assay was developed which permits rapid, quantitative assessment of migration across a membrane filter. The critical factor in the assay was the use of a 10 .mu.m thick polycarbonate membrane without the usual polyvinylpyrrolidone coating. Migrated neutrophils remain adherent to the uncoated membrane, whereas 20-50% fall off polyvinylpyrrolidone-coated membranes. A major advantage of the method is that neutrophil chemotaxis can be readily quantified, since the migrated cells adhere to the membrane surface and are in 1 optical plane for counting. A 25 mm .times. 80 mm membrane sheet was used in a 48-well microchemotaxis assembly, which requires only 20,000 neutrophils and 25 .mu.l of attractant per assay well. Neutrophil chemotaxis was complete within 10-20 min at 37.degree. C, with 20-30% of the cells migrating to N-formyl-methionyl-leucyl-phenylalanine and 40-50% migrating to C5a [a fragment of complement component 5].