In vivo imaging with cellular resolution of bone marrow cells transplanted into the ischemic brain of a mouse.
The aim of the study was to monitor in vivo and noninvasively the fate of single bone marrow cells (BMCs) transplanted into the ischemic brain of unirradiated mice. In vivo imaging was performed through a closed cranial window, throughout the 2 weeks following cell transplantation, using laser-scanning confocal fluorescence microscopy. The window was chronically implanted above the left parieto-occipital cortex in C57BL/6J adult mice. BMC (3 x 10(5) nucleated cells in 0.5 microL medium) from 5-week-old transgenic mice, ubiquitously expressing green fluorescent protein (GFP), was transplanted into the ipsilateral cortex 24 h after the induction of focal ischemia by coagulation of the left middle cerebral artery (n = 15). Three nonischemic mice served as controls. Repeated in vivo imaging, up to a depth of 200 microm, revealed that BMCs survived within the ischemic and peri-ischemic cortex, migrated significantly towards the lesion, proliferated and adopted a microglia-like morphology over 2 weeks. These results were confirmed using ex vivo imaging after appropriate immunocytochemical treatments. This study indicates that confocal fluorescence microscopy is a reliable and unique tool to repeatedly assess with cellular resolution the in vivo dynamic fate of fluorescent cells transplanted into a mouse brain. These results also provide the first in vivo findings on the fate of single BMCs transplanted into the ischemic brain of unirradiated mice.
Tran-Dinh
Alexy
A
Cardiovascular Research Centre, INSERM U 689, Université Paris 7, 10 Avenue de Verdun, 75010 Paris, France.
Dinh
Alexy Tran
AT
Kubis
Nathalie
N
Tomita
Yutaka
Y
Karaszewski
Bartosz
B
Calando
Yolande
Y
Oudina
Karim
K
Petite
Hervé
H
Seylaz
Jacques
J
Pinard
Elisabeth
E
D000818:Animals; D001854:Bone Marrow Cells / Q000473:pathology*; D016026:Bone Marrow Transplantation / Q000473:pathology*; D002545:Brain Ischemia / Q000473:pathology*; D002455:Cell Division / Q000502:physiology; D002465:Cell Movement / Q000502:physiology; D002470:Cell Survival / Q000502:physiology; D002540:Cerebral Cortex / Q000098:blood supply / Q000473:pathology; D049452:Green Fluorescent Proteins / Q000032:analysis; D007091:Image Processing, Computer-Assisted; D020244:Infarction, Middle Cerebral Artery / Q000473:pathology*; D047569:Laser Scanning Cytometry; D051379:Mice; D008810:Mice, Inbred C57BL; D017628:Microglia / Q000473:pathology
D049452:Green Fluorescent Proteins