Tumor Biology

, Volume 37, Issue 8, pp 11375–11384 | Cite as

A comprehensive characterization of cell cultures and xenografts derived from a human verrucous penile carcinoma

  • Juan J. Muñoz
  • Sandra A. Drigo
  • Hellen Kuasne
  • Rolando A. R. Villacis
  • Fabio A. Marchi
  • Maria A. C. Domingues
  • Ademar Lopes
  • Tiago G. Santos
  • Silvia R. Rogatto
Original Article


This study aimed to establish and characterize primary cell cultures and xenografts derived from penile carcinoma (PeCa) in order to provide experimental models for cellular processes and efficacy of new treatments. A verrucous squamous cell carcinoma (VSCC) was macrodissected, dissociated, and cultivated in KSFM/DF12 medium. Cell cultures were evaluated at passage 5 (P5) using migration and invasion assays and were serially propagated, in vivo, in BALB/c nude mice until passage 3 (X1–X3). Immunophenotypic characterization of cultures and xenografts was performed. Genomic (CytoScan HD, Affymetrix) and transcriptomic profiles (HTA 2.0 platform, Affymetrix) for VSCC, cell cultures, and xenografts were assessed. P5 cells were able to migrate, invade the Matrigel, and produce tumors in immunodeficient mice, demonstrating their malignant potential. The xenografts unexpectedly presented a sarcomatoid-like carcinoma phenotype. Genomic analysis revealed a high similarity between the VSCC and tumor-derived xenograft, confirming its xenograft origin. Interestingly, a subpopulation of P5 cells presented stem cell-related markers (CD44+CD24 and ALDH1high) and sphere-forming capacity, suggesting their potential xenograft origin. Cell cultures and xenografts retained the genomic alterations present in the parental tumor. Compared to VSCC, differentially expressed transcripts detected in all experimental conditions were associated with cellular morphology, movement, and metabolism and organization pathways. Malignant cell cultures and xenografts derived from a verrucous penile carcinoma were established and fully characterized. Nevertheless, xenograft PeCa models must be used with caution, taking into consideration the selection of specific cell populations and anatomical sites for cell/tumor implantation.


Penile neoplasm Primary cell cultures Neoplasms Experimental disease models Mice 



The authors are grateful for the assistance given by MSc. Rainer Marco Lopes Lapa and Dr Vilma R Martins for the scientific support with the xenograft model.


This study was supported by grants from the National Council of Technological and Scientific Development (CNPq 573589/08-9) and São Paulo Research Foundation (FAPESP 2009/52088-3, 2010/51601-6, and 2009/14027-2).

Compliance with ethical standards

Conflicts of interest


Supplementary material

13277_2016_4951_MOESM1_ESM.tif (945 kb)
Fig. S1 Schematic flow chart of the study design including the establishment and characterization of verrucous penile carcinoma cell cultures and xenografts. IHC: immunohistochemistry; P1-P5: cell culture passage from first to fifth (in vitro), X1, X2 and X3: xenograft in the first, second and third passages (in vivo). Immunophenotyping of xenograft cell cultures (X1, X2 and X3) was done (rectangle with dashed lines). (TIF 945 kb)
13277_2016_4951_MOESM2_ESM.doc (42 kb)
Table S1 Antibodies used for phenotypic characterization assays. (DOC 41 kb)
13277_2016_4951_MOESM3_ESM.doc (39 kb)
Table S2 Phenotypic characterization of cell cultures and xenografts. (DOC 39 kb)
13277_2016_4951_MOESM4_ESM.doc (106 kb)
Table S3 Genomic alterations detected in cell cultures (P1 and P5) and xenograft on passage 1 (X1) in comparison with the parental penile carcinoma (VSCC). (DOC 106 kb)
13277_2016_4951_MOESM5_ESM.doc (54 kb)
Table S4 Top 20 differentially expressed genes detected in cell cultures (P1 and P5) and xenografts (X1-X3) in comparison with the parental penile carcinoma (VSCC). (DOC 54 kb)


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Copyright information

© International Society of Oncology and BioMarkers (ISOBM) 2016

Authors and Affiliations

  • Juan J. Muñoz
    • 1
    • 2
  • Sandra A. Drigo
    • 3
  • Hellen Kuasne
    • 1
    • 3
  • Rolando A. R. Villacis
    • 1
  • Fabio A. Marchi
    • 1
  • Maria A. C. Domingues
    • 4
  • Ademar Lopes
    • 5
  • Tiago G. Santos
    • 1
  • Silvia R. Rogatto
    • 1
    • 3
  1. 1.International Research Center—CIPE, A. C. Camargo Cancer CenterSão PauloBrazil
  2. 2.Genetic Graduation Program, Institute of Biosciences, São Paulo State University—UNESPBotucatuBrazil
  3. 3.NeoGene Laboratory, Department of UrologyMedical School, São Paulo State University—UNESPBotucatuBrazil
  4. 4.Department of Pathology, Medical SchoolSão Paulo State University—UNESPBotucatuBrazil
  5. 5.Department of Pelvic Surgery, A. C. Camargo Cancer CenterSão PauloBrazil

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