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Immunophenotypic characterisation and cytogenetic analysis of mesenchymal stem cells from equine bone marrow and foal umbilical cords during in vitro culture
2016
Mazurkevych, Anatoliy | Malyuk, Mykola | Bezdieniezhnykh, Natalia | Starodub, Lyubov | Kharkevych, Yuriy | Brusko, Evgen | Gryzińska, Magdalena | Jakubczak, Andrzej
Introduction: The objective of the study was immunophenotypic and cytogenetic analysis of mesenchymal stem cells from equine bone marrow and foal umbilical cords during in vitro culture.Material and Methods: The mesenchymal stem cells were obtained from equine bone marrow of three horses and from foal umbilical cords of six foals. The cells were cultured in CO₂ incubators by standard procedure. Quantitative abnormalities of chromosomes, i.e. aneuploidy and polyploidy, and structural aberrations, i.e. breaks in chromosomes and chromatid, were taken into account during the study.Results: The results of cytogenetic analysis of equine bone marrow mesenchymal stem cells at the third and fourth passages indicated that the level of karyotype variability of these cells corresponded to the spontaneous level of karyotype variability typical of the peripheral blood lymphocytes of this species. Equine bone marrow contained several clones of stem cells that differed in the expression of specific nuclear markers characteristic of proliferating cells.Conclusion: Mesenchymal stem cells from foal umbilical cords during in vitro cultivation are characterised by quantitative abnormalities of the chromosomal apparatus.
Mostrar más [+] Menos [-]Establishment of an immortalized cell line and transplantable xenograft from a bronchioloalveolar lung carcinoma of a cat
2002
Grossman, Deborah A. | McNiel, Elizabeth A. | Hackett, Tim B. | Barsky, Sanford H.
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.
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