HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text Full Text (PDF) All Versions of this Article: 2006-0013v1 24/9/2045    most recent Alert me when this article is cited Alert me if a correction is posted Citation Map Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints/Permissions Citing Articles Citing Articles via Google Scholar Google Scholar Articles by Spangrude, G. J. Articles by Fleming, W. H. Search for Related Content PubMed PubMed Citation Articles by Spangrude, G. J. Articles by Fleming, W. H.

TECHNOLOGY DEVELOPMENT

Mouse Models of Hematopoietic Engraftment: Limitations of Transgenic Green Fluorescent Protein Strains and a High-Performance Liquid Chromatography Approach to Analysis of Erythroid Chimerism

^aDepartment of Medicine,
^bDepartment of Pathology, and
^cDepartment of Neurobiology and Anatomy, University of Utah School of Medicine, Salt Lake City, Utah, USA;
^dDepartment of Medicine, Division of Hematology and Medical Oncology, Oregon Health & Science University, Portland, Oregon, USA

Key Words. Erythropoiesis • Hematopoietic chimerism • Hematopoietic cell transplantation • Engraftment

Correspondence: Gerald J. Spangrude, Ph.D., University of Utah, Division of Hematology RM 4C416, 30 N 1900 East, Salt Lake City, Utah 84132-2408, USA. Telephone: 801-585-5544; Fax: 801-585-3778; e-mail: gspangrude{at}mac.com

Received on January 6, 2006; accepted for publication on May 2, 2006.

First published online in STEM CELLS EXPRESS  May 11, 2006.


Transgenic mouse strains ubiquitously expressing green fluorescent protein (GFP) have enabled investigators to develop in vivo transplant models that can detect donor contributions to many different tissues. However, most GFP transgenics lack expression of the reporter in the erythroid lineage. We evaluated expression of GFP in the bone marrow of the OsbY01 transgenic mouse (B6-GFP) in the context of CD71 and TER-119 expression and found that GFP fluorescence is lost prior to the basophilic erythroblast stage of development. However, platelets in B6-GFP mice were found to be uniformly positive for GFP. We therefore used the GFP transgenic model in combination with allelic variants of CD45 and the hemoglobin ß (Hbb) chain to develop a model system that allows all blood lineages to be followed in a mouse model of bone marrow transplantation (BMT). To detect Hbb variant molecules, we developed a new protocol based on high-performance liquid chromatography that is sensitive and precise, allowing rapid and quantitative analysis of erythroid chimerism. Platelet and leukocyte engraftment were detected by flow cytometry. BMT into sublethally irradiated (4 Gy) recipients demonstrated the failure of B6-GFP-derived cells to engraft relative to B6-CD45^a-derived cells, suggesting that an immune barrier may prevent efficient engraftment of the transgenic cells in a setting of minimal ablation. These results establish limitations in the use of transgenic GFP expression as a donor marker in transplantation models.