| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH |
|
|
|
|||||||
|
|||||||||
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
TECHNOLOGY DEVELOPMENT |
1 Section of Digestive Diseases, Department of Internal Medicine, Yale University School of Medicine, New Haven, CT 06520
2 Department of Laboratory Medicine, Yale University School of Medicine, New Haven, CT 06520
3 Department of Surgery, Children's Hospital of Philadelphia, University of Pennsylvania School of Medicine, Philadelphia, PA 19104
* To whom correspondence should be addressed. E-mail: Scott.swenson{at}yale.edu.
 |
Abstract |
|---|
The enhanced green fluorescent protein (GFP) reporter has been widely adopted for tracking cell lineage. Here we compare three transgenic mouse strains in which GFP is considered "ubiquitously expressed", with the GFP transgene under control of the chicken beta actin (CBA) or human ubiquitin C (UBC) promoter. We compared the expression of GFP using flow cytometry, direct tissue fluorescence, and immunostaining with multiple commercially available anti-GFP antibodies. Mice of CBA-GFP strain "1Osb" have strong but variegated expression of GFP in adult liver, kidney, small intestine, and blood. Mice of CBA-GFP strain "Y01" have the highest proportion of GFP-positive peripheral blood cells, yet limited GFP expression in liver, intestine, and kidney. UBC-GFP mice express GFP only weakly in solid organs and variably in blood.
Direct fluorescent detection of GFP in formalin-fixed, paraffin-embedded tissue sections was the simplest approach, but only useful in high-expressing strains and potentially subject to artifact due to tissue autofluorescence. Immunofluorescence using either primary goat or rabbit antibodies was much more sensitive, and allowed better discrimination of authentic signal from autofluorescence. Immunohistochemical staining was less sensitive than direct fluorescence or immunofluorescence, and was subject to false positive signal in the small intestine. In conclusion, there is considerable variability of expression within and between GFP transgenic strains. None of the tested strains gives truly ubiquitous GFP expression. A detailed analysis of GFP expression in one's tissues of interest must guide the choice of reporter mouse strain when GFP is used as a marker of cell lineage or donor origin.
Key Words. green fluorescent protein, GFP, lineage tracing, bone marrow transplantation, donor marker
This article has been cited by other articles:
|
|
 |
|
  D. S. Krause Bone Marrow-derived Lung Epithelial Cells Proceedings of the ATS, August 15, 2008; 5(6): 699 - 702. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
D. J. Weiss, J. K. Kolls, L. A. Ortiz, A. Panoskaltsis-Mortari, and D. J. Prockop Stem Cells and Cell Therapies in Lung Biology and Lung Diseases Proceedings of the ATS, July 15, 2008; 5(5): 637 - 667. [Full Text] [PDF]
|
|
|
|
 |
|
 E. S. Swenson, I. Guest, Z. Ilic, M. Mazzeo-Helgevold, P. Lizardi, C. Hardiman, S. Sell, and D. S. Krause Hepatocyte Nuclear Factor-1 as Marker of Epithelial Phenotype Reveals Marrow-Derived Hepatocytes, but Not Duct Cells, After Liver Injury in Mice Stem Cells, July 1, 2008; 26(7): 1768 - 1777. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 G.-Y. Chen, C. Chen, L. Wang, X. Chang, P. Zheng, and Y. Liu Cutting Edge: Broad Expression of the FoxP3 Locus in Epithelial Cells: A Caution against Early Interpretation of Fatal Inflammatory Diseases following In Vivo Depletion of FoxP3-Expressing Cells J. Immunol., April 15, 2008; 180(8): 5163 - 5166. [Abstract] [Full Text] [PDF]
|
|
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH |
| STEM CELLS | THE ONCOLOGIST | CME | ALPHAMED PRESS JOURNALS |
