Open Access

Establishment of primary cell cultures from canine mammary gland malignant tumours: a preliminary study

Patŕicia Petrouškova

Patŕicia Petrouškova

Department of Epizootiology, Parasitology and Protection of One Health, University of Veterinary Medicine and Pharmacy in KošiceKošice, Slovak Republic
Orcid profile
Search for this author on
Sciendo|Google Scholar
Petrouškova, Patŕicia
, 
Nikola Hudáková

Nikola Hudáková

Small Animal Clinic, University Veterinary Hospital, University of Veterinary Medicine and Pharmacy in KošiceKošice, Slovak Republic
Orcid profile
Search for this author on
Sciendo|Google Scholar
Hudáková, Nikola
, 
Viera Almášiová

Viera Almášiová

Department of Morphological Disciplines, University of Veterinary Medicine and Pharmacy in KošiceKošice, Slovak Republic
Orcid profile
Search for this author on
Sciendo|Google Scholar
Almášiová, Viera
, 
Alexandra Valenčáková

Alexandra Valenčáková

Small Animal Clinic, University Veterinary Hospital, University of Veterinary Medicine and Pharmacy in KošiceKošice, Slovak Republic
Orcid profile
Search for this author on
Sciendo|Google Scholar
Valenčáková, Alexandra
, 
L’ubica Horňáková

L’ubica Horňáková

Small Animal Clinic, University Veterinary Hospital, University of Veterinary Medicine and Pharmacy in KošiceKošice, Slovak Republic
Orcid profile
Search for this author on
Sciendo|Google Scholar
Horňáková, L’ubica
, 
Mykhailo Huniadi

Mykhailo Huniadi

Small Animal Clinic, University Veterinary Hospital, University of Veterinary Medicine and Pharmacy in KošiceKošice, Slovak Republic
Orcid profile
Search for this author on
Sciendo|Google Scholar
Huniadi, Mykhailo
 and 
Daša Čížková

Daša Čížková

  • Small Animal Clinic, University Veterinary Hospital, University of Veterinary Medicine and Pharmacy in KošiceKošice, Slovak Republic
  • Institute of Neuroimmunology, Slovak Academy of Sciences v.v.i.Slovak Republic
Orcid profile
Search for this author on
Sciendo|Google Scholar
Čížková, Daša
  
Mar 01, 2025
Establishment of primary cell cultures from canine mammary gland malignant tumours: a preliminary study's Cover Image
About this article
Previous Article
Next Article
Cite
Download Cover
Published Online: Mar 01, 2025
Page range: 159 - 168
Received: Jul 17, 2024
Accepted: Feb 10, 2025
DOI: https://doi.org/10.2478/jvetres-2025-0007
Keywords
© 2025 Patŕicia Petrouškova et al., published by Sciendo
This work is licensed under the Creative Commons Attribution 4.0 International License.

Fig. 1.

Microphotographs of mammary gland tumours. A – magnification 200×, scale bar 50 μm; B – magnification 400×, scale bar 20 μm
Microphotographs of mammary gland tumours. A – magnification 200×, scale bar 50 μm; B – magnification 400×, scale bar 20 μm

Fig. 2.

Primary cell cultures derived from mammary gland solid adenocarcinoma and carcinosarcoma. A and G – cellular debris and blood cells present directly after plating the cells on day in vitro (DIV) 0; B and H – cells before washing on DIV 1; C and I – cells after washing on DIV 1. Washing the cells effectively removed the majority of cellular debris and non-adherent cells; D and J – enlarged irregular and star-shaped cells on DIV 3; E and K – larger polygonal and irregularly shaped cells on DIV 5; F and L – cells with a range of morphologies from spindle-shaped to polygonal-shaped on DIV 7. Magnification 200×, scale bar 50 μm
Primary cell cultures derived from mammary gland solid adenocarcinoma and carcinosarcoma. A and G – cellular debris and blood cells present directly after plating the cells on day in vitro (DIV) 0; B and H – cells before washing on DIV 1; C and I – cells after washing on DIV 1. Washing the cells effectively removed the majority of cellular debris and non-adherent cells; D and J – enlarged irregular and star-shaped cells on DIV 3; E and K – larger polygonal and irregularly shaped cells on DIV 5; F and L – cells with a range of morphologies from spindle-shaped to polygonal-shaped on DIV 7. Magnification 200×, scale bar 50 μm

Fig. 3.

Immunofluorescence staining to show mucin 1 (MUC1), cytokeratins 8 and 18 (CK8/18) and Kiel 67 (Ki-67) expression in canine mammary gland tumour cells. A and B – solid adenocarcinoma-derived and carcinosarcoma-derived cells expressing MUC1; C – solid adenocarcinoma-derived cells showing higher expression of CK8/18 than carcinosarcoma-derived cells; D – carcinosarcoma-derived cells showing lower expression of CK8/18 than solid adenocarcinoma-derived cells; E – solid adenocarcinoma-derived cells showing lower expression of Ki-67 than carcinosarcoma-derived cells; F – carcinosarcoma-derived cells showing higher expression of Ki-67 than solid adenocarcinoma-derived cells; G and H – 400× magnification of details of E and F. Nuclei were stained with 4′,6-diamidino-2-phenylindole. Magnification 100×, scale bar 100 μm
Immunofluorescence staining to show mucin 1 (MUC1), cytokeratins 8 and 18 (CK8/18) and Kiel 67 (Ki-67) expression in canine mammary gland tumour cells. A and B – solid adenocarcinoma-derived and carcinosarcoma-derived cells expressing MUC1; C – solid adenocarcinoma-derived cells showing higher expression of CK8/18 than carcinosarcoma-derived cells; D – carcinosarcoma-derived cells showing lower expression of CK8/18 than solid adenocarcinoma-derived cells; E – solid adenocarcinoma-derived cells showing lower expression of Ki-67 than carcinosarcoma-derived cells; F – carcinosarcoma-derived cells showing higher expression of Ki-67 than solid adenocarcinoma-derived cells; G and H – 400× magnification of details of E and F. Nuclei were stained with 4′,6-diamidino-2-phenylindole. Magnification 100×, scale bar 100 μm

Fig. 4.

Immunocytochemistry controls verifying the specificity of primary antibody staining. A – non-cancerous cells isolated from canine mammary glands demonstrating weak positive staining for mucin 1 (MUC1) (I) and cytokeratins 8 and 18 (CK8/18) (II), and minimal Kiel 67 (Ki-67) staining (III). (IV) is a 400× magnification of details of (III); B – negative controls where primary antibodies (MUC1 (I), CK8/18 (II) and Ki-67 (III)) were excluded from the assay. Nuclei were stained with 4′,6-diamidino-2-phenylindole. Magnification ×100, scale bar 100 μm
Immunocytochemistry controls verifying the specificity of primary antibody staining. A – non-cancerous cells isolated from canine mammary glands demonstrating weak positive staining for mucin 1 (MUC1) (I) and cytokeratins 8 and 18 (CK8/18) (II), and minimal Kiel 67 (Ki-67) staining (III). (IV) is a 400× magnification of details of (III); B – negative controls where primary antibodies (MUC1 (I), CK8/18 (II) and Ki-67 (III)) were excluded from the assay. Nuclei were stained with 4′,6-diamidino-2-phenylindole. Magnification ×100, scale bar 100 μm

Specific primary and secondary antibodies used for immunocytochemistry

Primary antibodies
Marker Primary antibody Type, clone Host, isotype Reactivity Dilution Catalogue No. Supplier
mucin 1 MUC1 mAb, MUC1/955 mouse, IgG human, mouse 1 : 150 NBP2-44658 Novus Biologicals, Centennial, CO, USA
cytokeratin 8 cytokeratin 18 Cytokeratins 8/18 mAb, 5D3 mouse, IgG human, mouse 1 : 100 NB120-17139 Novus Biologicals
Ki-67 Anti-Ki-67 pAb, N/A rabbit, IgG human, mouse 1 : 300 ab15580 Abcam, Cambridge, UK
Language:
English
Publication timeframe:
4 times per year
Journal Subjects:
Life Sciences, Molecular Biology, Microbiology and Virology, Life Sciences, other, Medicine, Veterinary Medicine
Journal RSS Feed