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Establishment of an immortalized cell line and transplantable xenograft from a bronchioloalveolar lung carcinoma of a cat

Deborah A. GrossmanDepartment of Pathology, UCLA School of Medicine, Los Angeles, CA 90024.

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Elizabeth A. McNielDepartment of Radiological Health, College of Veterinary Medicine, Colorado State University, Fort Collins, CO 80523.

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Timothy B. HackettDepartment of Clinical Sciences, College of Veterinary Medicine, Colorado State University, Fort Collins, CO 80523.

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Sanford H. BarskyDepartment of Pathology, UCLA School of Medicine, Los Angeles, CA 90024.

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Abstract

Objective—To establish an immortalized cell line and transplantable xenograft of feline bronchioloalveolar lung carcinoma (BAC).

Sample Population—Pleural effusion from a 12-yearold Persian male cat with BAC.

Procedure—Tumor cells from the pleural effusion were grown in monolayer cell culture and injected into severe combined immunodeficient (SCID) mice to establish an immortalized cell line as well as a transplantable xenograft.

Results—Both the primary lung carcinoma, the derived cell line, and the transplantable xenograft had evidence of a type-II pneumocyte origin expressing lamellar bodies ultrastructurally and thyroid transcription factor-1 and surfactant immunocytochemically. All 3 also expressed nuclear p53 immunoreactivity. A metaphase spread of the cell line (SPARKY) probed with fluorescein-labeled genomic feline DNA gave evidence of its feline origin. Flow cytometric studies indicated aneuploidy with a DNA index of 1.6. An R-banded karyotype revealed a modal number of 66 including the feline Y chromosome. The cell line had a doubling time of 16 hours. The xenograft (SPARKY-X) reached a diameter of 1 cm in 3 weeks in SCID mice. Deoxyribonucleic acid fingerprint analysis revealed that SPARKY and SPARKY-X were novel and strongly matched each other, except for the murine component found in SPARKY-X. Interestingly, SPARKY-X manifested the characteristic lepidic growth pattern of pulmonic BAC.

Conclusions—Both the cell line and xenograft retained their autochthonous BAC phenotype, making them useful for the subsequent dissection of molecular abnormalities in feline BAC and in vitro screening of chemotherapeutic agents. (Am J Vet Res 2002; 63:1745–1753)

Abstract

Objective—To establish an immortalized cell line and transplantable xenograft of feline bronchioloalveolar lung carcinoma (BAC).

Sample Population—Pleural effusion from a 12-yearold Persian male cat with BAC.

Procedure—Tumor cells from the pleural effusion were grown in monolayer cell culture and injected into severe combined immunodeficient (SCID) mice to establish an immortalized cell line as well as a transplantable xenograft.

Results—Both the primary lung carcinoma, the derived cell line, and the transplantable xenograft had evidence of a type-II pneumocyte origin expressing lamellar bodies ultrastructurally and thyroid transcription factor-1 and surfactant immunocytochemically. All 3 also expressed nuclear p53 immunoreactivity. A metaphase spread of the cell line (SPARKY) probed with fluorescein-labeled genomic feline DNA gave evidence of its feline origin. Flow cytometric studies indicated aneuploidy with a DNA index of 1.6. An R-banded karyotype revealed a modal number of 66 including the feline Y chromosome. The cell line had a doubling time of 16 hours. The xenograft (SPARKY-X) reached a diameter of 1 cm in 3 weeks in SCID mice. Deoxyribonucleic acid fingerprint analysis revealed that SPARKY and SPARKY-X were novel and strongly matched each other, except for the murine component found in SPARKY-X. Interestingly, SPARKY-X manifested the characteristic lepidic growth pattern of pulmonic BAC.

Conclusions—Both the cell line and xenograft retained their autochthonous BAC phenotype, making them useful for the subsequent dissection of molecular abnormalities in feline BAC and in vitro screening of chemotherapeutic agents. (Am J Vet Res 2002; 63:1745–1753)