Migration and differentiation of nuclear fluorescence-labeled bone marrow stromal cells after transplantation into cerebral infarct and spinal cord injury in mice.
There is increasing evidence that bone marrow stromal cells (BMSC) have the potential to migrate into the injured neural tissue and to differentiate into the CNS cells, indicating the possibility of autograft transplantation therapy. The present study was aimed to clarify whether the mouse BMSC can migrate into the lesion and differentiate into the CNS cells when transplanted into the mice subjected to focal cerebral infarct or spinal cord injury. The BMSC were harvested from mice and characterized by flow cytometry. Then, the BMSC were labeled by bis-benzimide, a nuclear fluorescence dye, over 24 h, and were stereotactically transplanted into the brain or spinal cord of the mice. The cultured BMSC expressed low levels of CD45 and high levels of CD90 and Sca-1 on flow cytometry. A large number of grafted cells survived in the normal brain 4 weeks after transplantation, many of which were located close to the transplanted sites. They expressed the neuronal marker including NeuN, MAP2, and doublecortin on fluorescent immunohistochemistry. However, when the BMSC were transplanted into the ipsilateral striatum of the mice subjected to middle cerebral artery occlusion, many of the grafted cells migrated into the corpus callosum and injured cortex, and also expressed the neuronal markers 4 weeks after transplantation. In particular, NeuN was very useful to validate the differentiation of the grafted cells, because the marker was expressed in the nuclei and was overlapped with bis-benzimide. Similar results were obtained in the mice subjected to spinal cord injury. However, many of the transplanted BMSC expressed GFAP, an astrocytic protein, in injured spinal cord. The present results indicate that the mouse BMSC can migrate into the CNS lesion and differentiate into the neurons or astrocytes, and that bis-benzimide is a simple and useful marker to label the donor cells and to evaluate their migration and differentiation in the host neural tissues over a long period.
Lee
JangBo
J
Department of Neurosurgery, Hokkaido University Graduate School of Medicine, Sapporo, Japan.
Kuroda
Satoshi
S
Shichinohe
Hideo
H
Ikeda
Jun
J
Seki
Toshitaka
T
Hida
Kazutoshi
K
Tada
Mitsuhiro
M
Sawada
Ken-ichi
K
Iwasaki
Yoshinobu
Y
D000818:Animals; D006690:Bisbenzimidazole / Q000494:pharmacology; D001854:Bone Marrow Cells / Q000378:metabolism; D016026:Bone Marrow Transplantation; D002454:Cell Differentiation; D002465:Cell Movement; D002544:Cerebral Infarction / Q000628:therapy*; D004195:Disease Models, Animal; D005434:Flow Cytometry; D005456:Fluorescent Dyes / Q000494:pharmacology; D005904:Glial Fibrillary Acidic Protein / Q000096:biosynthesis; D006085:Graft Survival; D008297:Male; D051379:Mice; D008869:Microtubule-Associated Proteins / Q000096:biosynthesis; D009457:Neuroglia / Q000166:cytology; D009474:Neurons / Q000166:cytology; D013119:Spinal Cord Injuries / Q000628:therapy*; D013194:Staining and Labeling; D033581:Stem Cell Transplantation; D017154:Stromal Cells / Q000637:transplantation
D005456:Fluorescent Dyes; D005904:Glial Fibrillary Acidic Protein; D008869:Microtubule-Associated Proteins; D006690:Bisbenzimidazole