title: Detection of chromosomal changes in chronic lymphocytic leukemia using classical cytogenetic methods and FISH: application of rich mitogen mixtures for lymphocyte cultures.
abstract: One of the research methods of prognostic value in chronic lymphocytic leukemia (CLL) is cytogenetic analysis. This method requires the presence of appropriate B-cell mitogens in cultures in order to obtain a high mitotic index. The aim of our research was to determine the most effective methods of in vitro B-cell stimulation to maximize the number of metaphases from peripheral blood cells of patients with CLL for classical cytogenetic examination, and then to correlate the results with those obtained using fluorescence in situ hybridization (FISH). The study group involved 50 consecutive patients with CLL. Cell cultures were maintained with the basic composition of culture medium and addition of respective stimulators. We used the following stimulators: Pokeweed Mitogen (PWM), 12-O-tetradecanoylphorbol 13-acetate (TPA), ionophore, lipopolysaccharide (LPS), and CpG-oligonucleotide DSP30. We received the highest mitotic index when using the mixture of PWM+TPA+I+DSP30. With classical cytogenetic tests using banding techniques, numerical and structural aberrations of chromosomes were detected in 46 patients, and no change was found in only four patients. Test results clearly confirmed the legitimacy of using cell cultures enriched with the mixture of cell stimulators and combining classical cytogenetic techniques with the FISH technique in later patient diagnosing.
authors.affiliation: Department of Cancer Genetics, Medical University of Lublin, Lublin, Poland.
authors.forename: Dorota
authors.initials: D
authors.lastname: Koczkodaj
authors.affiliation: Department of Cancer Genetics, Medical University of Lublin, Lublin, Poland.
authors.forename: Sylwia
authors.initials: S
authors.lastname: Popek
authors.affiliation: Department of Cancer Genetics, Medical University of Lublin, Lublin, Poland.
authors.forename: Szymon
authors.initials: S
authors.lastname: Zmorzyński
authors.affiliation: Department of Hematooncology and Bone Marrow Transplantation, Medical University of Lublin, Lublin, Poland.
authors.forename: Ewa
authors.initials: E
authors.lastname: Wąsik-Szczepanek
authors.affiliation: Department of Cancer Genetics, Medical University of Lublin, Lublin, Poland.
authors.forename: Agata A
authors.initials: AA
authors.lastname: Filip
mesh_terms: D000368:Aged; D000369:Aged, 80 and over; D002478:Cells, Cultured; D002869:Chromosome Aberrations; D020732:Cytogenetic Analysis / Q000379:methods*; D005260:Female; D006801:Humans; D017404:In Situ Hybridization, Fluorescence / Q000379:methods*; D015451:Leukemia, Lymphocytic, Chronic, B-Cell / Q000235:genetics*; D008214:Lymphocytes / Q000187:drug effects / Q000378:metabolism*; D008297:Male; D008677:Metaphase / Q000187:drug effects; D008875:Middle Aged; D008934:Mitogens / Q000494:pharmacology*; D008940:Mitotic Index
chemical_list: D008934:Mitogens
keywords: Cell Proliferation; Hematologic Diseases; Leukemia