lacZ transduction of 9L rat tumor cells without cloning.
Several laboratories have introduced the lacZ gene into 9L cells to improve the usefulness of this already popular rat brain tumor model. However, these laboratories were not concerned about possible changes in the phenotypic characteristics of the 9L cell line which can be induced by the selection of lacZ-expressing clones. Here, we describe a method for introducing the lacZ gene into 9L cells without selective cloning. The 9L parent cells (passaged the same number of times) and 9L/lacZ cells were compared in a number of tests and found to have the same phenotype. Specifically, they had the same sensitivity to radiation from external gamma or internal beta radiation, the same growth rates with or without frequent media changes and the same patterns of growth in rat brain. We demonstrated that the 9L/lacZ cells could be sorted from dissociated tumors by flow cytometry and the percentage of nonmalignant versus malignant cells determined. These percentages were variable from rat to rat. The colony-forming efficiency could be determined on the basis of whole tumor or, by using the percent of lacZ-positive cells, on the basis of malignant cells in a tumor. These novel approaches should render the 9L tumor model even more useful.
Gabryel-Grudziak
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Department of Radiation Oncology, University of Pittsburgh School of Medicine, PA 15261, USA.
Boggs
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Redgate
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Deutsch
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D000818:Animals; D001921:Brain / Q000201:enzymology / Q000473:pathology; D001932:Brain Neoplasms / Q000201:enzymology / Q000235:genetics* / Q000473:pathology; D003001:Cloning, Molecular; D005434:Flow Cytometry; D005822:Genetic Vectors; D018316:Gliosarcoma / Q000201:enzymology / Q000235:genetics* / Q000473:pathology; D007763:Lac Operon / Q000235:genetics*; D010641:Phenotype; D051381:Rats; D011916:Rats, Inbred F344; D014161:Transduction, Genetic / Q000235:genetics*; D014407:Tumor Cells, Cultured / Q000473:pathology / Q000637:transplantation; D001616:beta-Galactosidase / Q000378:metabolism
D001616:beta-Galactosidase