TY - JOUR
T1 - 3D-confocal structural analysis of bone marrow-derived renal tubular cells during renal ischemia/reperfusion injury
AU - Toyokawa, Hideyoshi
AU - Nakao, Atsunori
AU - Stolz, Donna B.
AU - Romanosky, Anna J.
AU - Nalesnik, Michael A.
AU - Neto, Joao Seda
AU - Kaizu, Takashi
AU - Demetris, Anthony J.
AU - Murase, Noriko
N1 - Funding Information:
We thank Mark Ross, Jason Devlin and Stuart Shand for excellent support in 3D imaging, Mike Tabacek and Lisa Chedwick for technical support, Masaru Okabe for providing EGFP rats, and Carla Forsythe for the preparation and organization of manuscript. This work was supported by NIH Grants DK54232, CA76541 and DK49615.
Copyright:
Copyright 2008 Elsevier B.V., All rights reserved.
PY - 2006/1
Y1 - 2006/1
N2 - Bone marrow cells (BMC) have been shown to migrate into injured sites for parenchymal repair. However, the extent of BMC involvement is controversial. To determine whether and to what extent BMC contribute to renal parenchymal repair, we employed three-dimensional (3D) fluorescent confocal microscopy/video in renal warm and cold ischemia/reperfusion (I/R) injury using enhanced green fluorescent protein transgenic rats and their radiation chimeras. After induction of renal warm I/R injury in chimeras, BM-derived renal tubular cells were found in 2D microscopy as isolated single cells or clusters of 2-3 cells. Likewise, cold I/R injury resulted in host-derived tubular cells with frequencies ≃0.2%. However, stringent confocal microscopic analysis and 3D image construction revealed that BM-derived tubules identified in 2D images were frequently artifacts of overlapping cells separately stained with different markers. The actual frequency in 3D analysis was approximately one-fourth of that seen in 2D analysis. 3D confocal imaging precisely detected BM-derived tubular epithelial cells and could be useful to study BMC contribution to tissue repair.
AB - Bone marrow cells (BMC) have been shown to migrate into injured sites for parenchymal repair. However, the extent of BMC involvement is controversial. To determine whether and to what extent BMC contribute to renal parenchymal repair, we employed three-dimensional (3D) fluorescent confocal microscopy/video in renal warm and cold ischemia/reperfusion (I/R) injury using enhanced green fluorescent protein transgenic rats and their radiation chimeras. After induction of renal warm I/R injury in chimeras, BM-derived renal tubular cells were found in 2D microscopy as isolated single cells or clusters of 2-3 cells. Likewise, cold I/R injury resulted in host-derived tubular cells with frequencies ≃0.2%. However, stringent confocal microscopic analysis and 3D image construction revealed that BM-derived tubules identified in 2D images were frequently artifacts of overlapping cells separately stained with different markers. The actual frequency in 3D analysis was approximately one-fourth of that seen in 2D analysis. 3D confocal imaging precisely detected BM-derived tubular epithelial cells and could be useful to study BMC contribution to tissue repair.
KW - Bone marrow-derived cells
KW - GFP
KW - Ischemia/reperfusion injury
KW - Kidney transplantation
KW - Three dimension
KW - Tissue regeneration
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U2 - 10.1038/labinvest.3700363
DO - 10.1038/labinvest.3700363
M3 - Article
C2 - 16258520
AN - SCOPUS:32844472262
VL - 86
SP - 72
EP - 82
JO - Laboratory Investigation
JF - Laboratory Investigation
SN - 0023-6837
IS - 1
ER -