Isolation, establishment, and characterization of ex vivo equine melanoma cell cultures
| Autor: | Nadine Metzger, Sarah W. Kamau Chapman, Brigitte von Rechenberg, Michael O. Hottiger, Paula Grest, Jörg A Auer, Karsten Feige |
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| Přispěvatelé: | University of Zurich, Hottiger, Michael O |
| Rok vydání: | 2009 |
| Předmět: |
Pathology
medicine.medical_specialty Blotting Western 10184 Institute of Veterinary Pathology Cell Separation Biology Metastasis 1309 Developmental Biology 1307 Cell Biology Antigen Antigens Neoplasm Proliferating Cell Nuclear Antigen Dispase medicine Tumor Cells Cultured Animals Horses neoplasms Melanoma Melanoma-associated antigen Microscopy Confocal 11077 Center for Applied Biotechnology and Molecular Medicine Staining and Labeling Cell Biology General Medicine medicine.disease Flow Cytometry 10226 Department of Molecular Mechanisms of Disease Cell culture Immunology 570 Life sciences biology 10090 Equine Department Stem cell Ex vivo Developmental Biology |
| DOI: | 10.5167/uzh-9500 |
| Popis: | Gray horses spontaneously develop metastatic melanomas that resemble human disease, and this is often accompanied with metastasis to other organs. Unlike in other species, the establishment of primary equine melanoma cultures that could be used to develop new therapeutic approaches has remained a major challenge. The purpose of the study was to develop a protocol for routine isolation and cultivation of primary equine melanocytes. Melanoma tissues were excised from 13 horses under local anesthesia, mainly from the perianal area. The melanoma cells were isolated from the melanoma tissue by serial enzymatic digestion using dispase and collagenase. Out of the 13 excised melanomas, cell cultures from eight melanomas were established, which corresponded to a success rate 62%. These cells showed different degrees of melanin pigmentation. Characterization of these cells using confocal microscopy, FACs analysis and western blotting showed that they expressed melanoma-associated antigens; Melan-A, MAGE-1, and MAGE-3, and PCNA expression was higher in fast-proliferating isolates. The protocol we developed and established proved successful for routine isolation and cultivation of primary equine melanoma cells. This method provided a large number of primary equine melanoma cells that could be used to study new therapeutic approaches for treatment of equine melanomas. |
| Databáze: | OpenAIRE |
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