Cryopreserved cytoplasts from human neutrophils migrate across monolayers of human endothelial cells in response to a chemoattractant gradient.
We have measured the capacity of two types of granule-poor anucleate cytoplasmic fragments (cytoplasts) from human polymorphonuclear leukocytes (PMN) to migrate across the barrier imposed by monolayers of human umbilical vein endothelial cells (EC) with and without chemotactic stimulation by fMLP. Cytoplasts were made by brief heating of PMN attached to surfaces (CKP) or by discontinuous gradient centrifugation (U-CYT). In the absence of chemoattractant, both types of cytoplast adhered poorly to endothelial cell monolayers, as did unstimulated intact PMN from which the cytoplasts were derived. In the presence of a transendothelial chemoattractant gradient both types of cytoplast exhibited a marked increase in adherence to, and migration across, endothelial monolayers; CKP did so to the same extent as chemoattractant-stimulated intact PMN. Since these motile cytoplasts are markedly deficient in most cytoplasmic organelles they may serve as useful tools for the dissection of cellular mechanisms that mediate PMN migration across endothelia.
Department of Medicine, College of Physicians and Surgeons, Columbia University, New York, NY 10032.
A J
AJ
Huang
S C
SC
Silverstein
S E
SE
Malawista
D002465:Cell Movement; D002634:Chemotaxis, Leukocyte; D004730:Endothelium, Vascular / Q000502:physiology; D005615:Freezing; D006801:Humans; D066298:In Vitro Techniques; D008854:Microscopy, Electron; D009240:N-Formylmethionine Leucyl-Phenylalanine / Q000494:pharmacology; D009504:Neutrophils / Q000502:physiology* / Q000648:ultrastructure; D011309:Preservation, Biological; D015398:Signal Transduction
D009240:N-Formylmethionine Leucyl-Phenylalanine