Porcine repeat element DNA: in situ detection of xenotransplanted cells.
Using a digoxygenin-labelled DNA probe derived from the porcine repeat element PRE-1, we have developed a protocol for the detection of transplanted porcine islets and hepatocytes against a background of murine host tissue. Analysis of this probe by Southern blotting indicated that PRE-1 hybridizes to pig genomic DNA but not to human or mouse DNA. On tissue sections, hybridizing probe was detected using alkaline phosphatase-conjugated antidigoxygenin antibody visualized with 5-bromo-4-chloro-3-indolyl-phosphate/4-nitro-blue tetrazolium chloride (BCIP/NBT) substrate. We have demonstrated sensitive and highly specific staining of porcine nuclei in fixed, paraffin embedded tissue sections, and have applied the technique to detect porcine pancreatic islets and hepatocytes transplanted into murine kidney and spleen. Application of this technique include detection of transplanted cells or organs across the variety of xenogeneic barriers.
Oettinger
H F
HF
Department of Molecular and Cellular Biology, Diacrin, Inc., Charlestown, MA 02129, USA.
Rodrigue-Way
A
A
Bousquet
J J
JJ
Edge
A S
AS
D000818:Animals; D017690:Cell Transplantation / Q000379:methods*; D015342:DNA Probes; D006801:Humans; D017403:In Situ Hybridization; D016381:Islets of Langerhans Transplantation / Q000473:pathology; D007668:Kidney / Q000378:metabolism / Q000473:pathology; D016031:Liver Transplantation; D051379:Mice; D008807:Mice, Inbred BALB C; D012091:Repetitive Sequences, Nucleic Acid; D013154:Spleen / Q000378:metabolism / Q000473:pathology; D013552:Swine / Q000235:genetics; D014183:Transplantation, Heterologous
D015342:DNA Probes