M.S. (Master of Science)
Department of Biological Sciences
Human cytogenetics; Flow cytometry; Lymphocytes
This thesis describes the flow cytometric analysis of several normal and transformed human cell lines and normal lymphocytes. Three methods of isolation and staining of chromosomes for flow cytometric analysis were investigated and the Aten method was found to be superior. Many different instrumentation variables were explored and optimal settings for clear chromosome analysis and resolution were established. Row karyotypic analysis of chromosomes from normal human cell lines produced normal chromosome distributions with no marked abnormalities. Normal phytohemagglutinin-stimulated lymphocytes also produced normal chromosome histograms, and it was found that culture for 5 days was optimal for chromosome resolution. The WI-38 SV cell line was found to be karyotypically unstable, and thus histograms were not obtainable with this method of analysis. Two of the cell lines, GM0637D and GM04312A, were found by flow karyotype analysis to be contaminated with Chinese hamster ovary cells. In addition, a series of colon carcinoma cell lines were analyzed by DNA cell cycle analysis and flow karyotyping. In spite of marked changes in cell cycle kinetics, no correlation between aneuploidy and abnormal flow karyotype was found. A large hamster chromosome (number 1) and a small hamster chromosome (number 8) were each sorted from a chromosome suspension isolated from the Chinese hamster ovary cell line. Analysis of the sorted chromosomes resulted in a sort purity of 67 and 69% respectively.
Aller, Cynthia Barbara Timlick, "Flow cytometric analysis of normal and transformed human cell lines and normal lymphocytes : karyotyping, sorting of chromosomes, and cell cycle analysis" (1989). Graduate Research Theses & Dissertations. 3154.
x, 122 pages
Northern Illinois University
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