Dual-label centromere and telomere FISH identifies human, rat, and mouse cell contribution to Multispecies recombinant urogenital sinus xenografts.
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.
Vander Griend
Donald J
DJ
Chemical Therapeutics Program, The Sidney Kimmel Comprehensive Cancer Center at Johns Hopkins, Baltimore, Maryland 21231, USA.
Konishi
Yuko
Y
De Marzo
Angelo M
AM
Isaacs
John T
JT
Meeker
Alan K
AK
D000818:Animals; D015415:Biomarkers; D001706:Biopsy; D002469:Cell Separation / Q000379:methods; D002503:Centromere / Q000235:genetics; D004848:Epithelium / Q000196:embryology / Q000637:transplantation; D005260:Female; D006801:Humans; D016867:Immunocompromised Host; D017404:In Situ Hybridization, Fluorescence / Q000379:methods*; D008297:Male; D008648:Mesoderm / Q000166:cytology / Q000196:embryology; D051379:Mice; D009924:Organ Culture Techniques / Q000379:methods; D016612:Paraffin Embedding; D011247:Pregnancy; D011467:Prostate / Q000166:cytology* / Q000196:embryology / Q000637:transplantation*; D051381:Rats; D017207:Rats, Sprague-Dawley; D011994:Recombinant Proteins / Q000235:genetics; D013045:Species Specificity; D016615:Telomere / Q000235:genetics; D014183:Transplantation, Heterologous / Q000379:methods*
D015415:Biomarkers; D011994:Recombinant Proteins