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Home / Equine Research Bank / In Vitro Cell Dev Biol Anim / Isolation, establishment, and characterization of ex vivo eq...
In vitro cellular & developmental biology. Animal2008; 45(3-4); 152-162; doi: 10.1007/s11626-008-9156-3

Isolation, establishment, and characterization of ex vivo equine melanoma cell cultures.

Abstract: 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.
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Publication Date: 2008-12-05 PubMed ID: 19057970DOI: 10.1007/s11626-008-9156-3Google Scholar: Lookup
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  • Non-U.S. Gov't

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

The research article focuses on the development of a protocol for the extraction and cultivation of melanoma cells from horses for the purpose of studying new treatment approaches. Primary equine melanoma cells were successfully isolated and cultivated from tissues excised from various horses, which expressed distinct melanoma-associated antigens and showed varying rates of pigmentation and growth.

Protocol Development for Equine Melanoma Cell Cultivation

  • The study aimed to tackle the problem of difficulty in establishing primary equine melanoma cultures, a critical step in devising new therapeutic strategies.
  • The solution was the development of a systematic method for routine isolation and cultivation of primary equine melanocytes.
  • The study involved excising melanoma tissues from 13 horses, primarily from the perianal area under local anesthesia.
  • The melanoma cells were then isolated from the tissue by enzymatic digestion using dispase and collagenase in a series.

Results and Characteristics of Equine Melanoma Cell Cultures

  • Of the 13 melanomas excised, cell cultures were established from eight, corresponding to a success rate of 62%.
  • These cells exhibited varying levels of melanin pigmentation, demonstrating their individual characteristics.
  • Confocal microscopy, FACs analysis, and western blotting techniques were used to test these cells.
  • The results showed the cells expressed melanoma-associated antigens; Melan-A, MAGE-1, and MAGE-3.
  • Particularly, cells that proliferated swiftly showed elevated expression of PCNA (Proliferating Cell Nuclear Antigen), a key protein involved in cellular replication.

Implications and Conclusion

  • The study’s success in developing and establishing this protocol may pave the way for wider, more practicable isolation and cultivation of primary equine melanoma cells.
  • This capability may further enable the scientific community to study and formulate new treatment approaches for this particular type of cancer.
  • Through the established protocol, it is possible to obtain a rich source of melanoma cells from horses, providing a significant advance in equine melanoma research.

Cite This Article

APA
Chapman SW, Metzger N, Grest P, Feige K, von Rechenberg B, Auer JA, Hottiger MO. (2008). Isolation, establishment, and characterization of ex vivo equine melanoma cell cultures. In Vitro Cell Dev Biol Anim, 45(3-4), 152-162. https://doi.org/10.1007/s11626-008-9156-3

Publication

In vitro cellular & developmental biology. Animal
ISSN: 1543-706X
NlmUniqueID: 9418515
Country: Germany
Language: English
Volume: 45
Issue: 3-4
Pages: 152-162

Researcher Affiliations

Chapman, Sarah W Kamau
  • Institute of Veterinary Biochemistry and Molecular Biology, Vetsuisse Faculty, University of Zurich, Winterthurerstr. 190, CH-8057, Zurich, Switzerland.
Metzger, Nadine
    Grest, Paula
      Feige, Karsten
        von Rechenberg, Brigitte
          Auer, Jörg A
            Hottiger, Michael O

              MeSH Terms

              • Animals
              • Antigens, Neoplasm / metabolism
              • Blotting, Western
              • Cell Separation / methods
              • Flow Cytometry
              • Horses
              • Melanoma / pathology
              • Microscopy, Confocal
              • Proliferating Cell Nuclear Antigen / metabolism
              • Staining and Labeling
              • Tumor Cells, Cultured

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              Citations

              This article has been cited 3 times.
              1. Piotrowska A, Zaucha R, Król O, Żmijewski MA. Vitamin D Modulates the Response of Patient-Derived Metastatic Melanoma Cells to Anticancer Drugs. Int J Mol Sci 2023 Apr 28;24(9).
                doi: 10.3390/ijms24098037pubmed: 37175742google scholar: lookup
              2. Seltenhammer MH, Sundström E, Meisslitzer-Ruppitsch C, Cejka P, Kosiuk J, Neumüller J, Almeder M, Majdic O, Steinberger P, Losert UM, Stöckl J, Andersson L, Sölkner J, Vetterlein M, Golovko A. Establishment and characterization of a primary and a metastatic melanoma cell line from Grey horses. In Vitro Cell Dev Biol Anim 2014 Jan;50(1):56-65.
                doi: 10.1007/s11626-013-9678-1pubmed: 23982913google scholar: lookup
              3. Sundström E, Imsland F, Mikko S, Wade C, Sigurdsson S, Pielberg GR, Golovko A, Curik I, Seltenhammer MH, Sölkner J, Lindblad-Toh K, Andersson L. Copy number expansion of the STX17 duplication in melanoma tissue from Grey horses. BMC Genomics 2012 Aug 2;13:365.
                doi: 10.1186/1471-2164-13-365pubmed: 22857264google scholar: lookup
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