Dual-label centromere and telomere FISH identifies human, rat, and mouse cell contribution to Multispecies recombinant urogenital sinus xenografts

Prostate. 2009 Oct 1;69(14):1557-64. doi: 10.1002/pros.21001.

Abstract

Background: Recombinant xenografts of human cells growing in immunocompromised rodents are widely used for studying stem cell biology, tumor biology, and epithelial to mesenchyme transitions. Of critical importance is the correct interpretation of the cellular composition of such xenografts.

Methods: Here we present a rapid and robust method employing protein nucleic acid (PNA) FISH probes to dual-label centromeres and telomeres (Cen/Tel FISH). Such labeling allows unambiguous discrimination between human, mouse, and rat cells in paraffin-embedded tissue sections, providing significant advantages over current methods used to discern human versus rodent cell types.

Results: Using an in vivo prostatic developmental system where rat embryonic urogenital sinus mesenchyme is recombined with human prostate epithelial organoids and grown in an immunocompromised mouse, Cen/Tel FISH documents that all three species contribute to the development of glandular structures.

Conclusions: The method is an indispensable tool to analyze xenograft/host interactions and prevent misinterpretation of data using tissue recombination approaches.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Animals
  • Biomarkers
  • Biopsy
  • Cell Separation / methods
  • Centromere / genetics
  • Epithelium / embryology
  • Epithelium / transplantation
  • Female
  • Humans
  • Immunocompromised Host
  • In Situ Hybridization, Fluorescence / methods*
  • Male
  • Mesoderm / cytology
  • Mesoderm / embryology
  • Mice
  • Organ Culture Techniques / methods
  • Paraffin Embedding
  • Pregnancy
  • Prostate / cytology*
  • Prostate / embryology
  • Prostate / transplantation*
  • Rats
  • Rats, Sprague-Dawley
  • Recombinant Proteins / genetics
  • Species Specificity
  • Telomere / genetics
  • Transplantation, Heterologous / methods*

Substances

  • Biomarkers
  • Recombinant Proteins