Laser-assisted microdissection and short tandem repeat PCR for the investigation of graft chimerism after solid organ transplantation.
The detection of donor-derived cells in the blood and tissues of graft recipients after solid organ transplantation is a readily observed phenomenon called microchimerism. Yet very little is known about the persistence and integration of recipient-derived cells in the transplanted organ, indicating a form of intragraft chimerism. To further study this phenomenon and its possible influence on graft acceptance or rejection, we developed the following novel approach. Immunohistochemically labeled cells were isolated by means of laser-based microdissection and subsequent laser pressure catapulting from paraffine-embedded posttransplantation biopsies. The following use of a highly sensitive PCR assay analyzing one polymorphic short tandem repeat (STR) marker enabled us to clearly identify the genotypes in samples containing as little as 10 isolated cells. The combination of laser-based microdissection and STR-PCR thus provides a powerful tool for the genotyping of even very few cells isolated from routinely processed biopsies after solid organ transplantation.
Institute of Pathology, Medizinische Hochschule Hannover, Germany. kleeberger.wolfram@mh-hannover.de
W
W
Kleeberger
T
T
Rothämel
S
S
Glöckner
U
U
Lehmann
H
H
Kreipe
D002469:Cell Separation / Q000295:instrumentation / Q000379:methods; D004210:Dissection / Q000295:instrumentation / Q000379:methods; D004730:Endothelium, Vascular / Q000473:pathology; D016027:Heart Transplantation / Q000473:pathology; D022781:Hepatocytes / Q000737:chemistry / Q000378:metabolism / Q000473:pathology; D006801:Humans; D007834:Lasers; D008099:Liver / Q000473:pathology; D016031:Liver Transplantation / Q000473:pathology; D009206:Myocardium / Q000473:pathology; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D020080:Tandem Repeat Sequences / Q000235:genetics*; D018183:Transplantation Chimera / Q000235:genetics*