Use of the Matrigel-based assay to measure the invasiveness of leukemic cells.
The reconstituted basement membrane (Matrigel)-based assay was used to quantify the invasive potential of hematopoietic cells including cultured human leukemic cells (KG-1, K-562, HEL, HL-60, and U-937), normal bone marrow (BM) cells, and normal polymorphonuclear leukocytes (PMNL). We found that (i) in contrast to 6- to 72-hour incubation periods typically used in assays with solid tumor cells, most of the invasive cell populations tested here required only 2 to 4 hours to cross the Matrigel layer; (ii) unlike that of PMNL, whose invasiveness was stimulated by the addition of FMLP, the invasive rate of cultured leukemic cells was not affected by this chemoattractant; (iii) the rate of invasion was inversely proportional to the Matrigel concentration per filter but varied with the Matrigel batch used; (iv) the most consistent results were obtained when 2.5 to 4 x 10(5) cells were added to the top portion of the blind well; and (v) of all leukemic cells tested, the least differentiated myeloblastic KG-1 cells exhibited the highest invasive potential, which was comparable to that of normal PMNL. We conclude that the Matrigel-based assay can be used as a model system in studies of mechanisms regulating movement of hematopoietic cells across basement membrane barriers.
Department of Medicine, Cross Cancer Institute, Edmonton, Alberta, Canada.
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Janiak
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Hashmi
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Janowska-Wieczorek
D001485:Basement Membrane; D001854:Bone Marrow Cells; D002452:Cell Count; D002465:Cell Movement; D003094:Collagen / Q000494:pharmacology; D004338:Drug Combinations; D005312:Fetal Blood; D006412:Hematopoietic Stem Cells / Q000473:pathology; D006801:Humans; D007797:Laminin / Q000494:pharmacology; D007938:Leukemia / Q000473:pathology*; D009240:N-Formylmethionine Leucyl-Phenylalanine / Q000494:pharmacology; D009361:Neoplasm Invasiveness; D009504:Neutrophils / Q000166:cytology; D011509:Proteoglycans / Q000494:pharmacology; D014407:Tumor Cells, Cultured
D004338:Drug Combinations; D007797:Laminin; D011509:Proteoglycans; C059485:matrigel; D009240:N-Formylmethionine Leucyl-Phenylalanine; D003094:Collagen