Format

Send to

Choose Destination
See comment in PubMed Commons below
Stem Cells. 2005 Jun-Jul;23(6):828-33.

Duplex polymerase chain reaction quantification of human cells in a murine background.

Author information

1
Laboratory of Molecular Medicine, Interdisiplinary Centre for Clinical Research, University of Leipzig Faculty of Medicine, Germany.

Abstract

Studies of the regenerative potential of human stem cells commonly involve their transplantation into immune-deficient mice or in vitro coculture with mouse cells. The optimal use of such models requires the detection and quantification of relatively low numbers of human cells in a murine background. We report here a duplex polymerase chain reaction (PCR) approach involving the coamplification of human-and mouse-specific repetitive sequences. The determination of product ratios compensates against variations in sample quality and enables quantitation from >50% down to 0.01% human-in-mouse from a single reaction. Product ratios are determined by standard electrophoresis of end-stage PCR reactions followed by image analysis techniques using freely available software, with no requirement for real-time PCR. The approach has been used to analyze tissue from mice transplanted with human cells and cocultures between differentiating mouse embryonal stem cells and human umbilical cord blood cells.

PMID:
15917478
DOI:
10.1634/stemcells.2004-0206
[Indexed for MEDLINE]
Free full text
PubMed Commons home

PubMed Commons

0 comments
How to join PubMed Commons

    Supplemental Content

    Full text links

    Icon for Wiley
    Loading ...
    Support Center