[Ferumoxide labeled Flk1+ CD31- CD34- human bone marrow mesenchymal stem cells and its in vivo tracing in the brains of Macaca Fascicularis].
OBJECTIVE
To explore the method for labeling Flk1+ CD31- CD34- human bone marrow mesenchymal stem cells (hBMSCs) with ferumoxide-PLL and evaluate the feasibility of its tracing after transplantation into the brains of Macaca Fascicularis.


METHODS
The hBMSCs were incubated with ferumoxide-PLL. Trypan blue staining, Prussian blue staining, and transmission electron microscope were performed to show intracellular iron, marking efficiency, and the vigor of the labeled cells. After the hBMSCs were transplanted into the brains of cynomolgus monkeys by stereotaxis, magnetic resonance imaging (MRI) was performed to trace the cells in vivo. Cell survival and differentiation were studied with immunohistochemistry, Prussian blue staining, and HE staining.


RESULTS
The marking efficiency of the ferumoxide-PLL was 96%. Iron particles were found intracytoplasmic of the hBMSCs by Prussian blue staining and transmission electron microscopy. The relaxation rates of labeled cells in MRI were 4.4 and 4.2 times higher than those of the unlabeled cells. Hypointensity area was found by MRI three weeks after transplantation. Many hBMSCs and new vessels were found in the transplantation zone by pathological and immunofluorescence methods.


CONCLUSIONS
Ferumoxide-PLL can effectively label hBMSCs and thus increase its contrast in MRI results. The cells can survive in the brains of cynomolgus monkeys. The labeled hBMSCs can be traced in vivo by MRI.
Feng
Ming
M
Department of Neurosurgery, PUMC Hospital, CAMS and PUMC, Beijing 100730, China.
Wang
Ren-Zhi
RZ
Zhu
Hua
H
Zhang
Nan
N
Wang
Chang-Jun
CJ
Wei
Jun-Ji
JJ
Lu
Shan
S
Li
Qin
Q
Yin
Xiao-Ming
XM
Han
Qin
Q
Ma
Wen-Bin
WB
Qin
Chuang
C
Zhao
Chun-Hua
CH
An
Yi-Hua
YH
Kong
Yan-Guo
YG
D000818:Animals; D018952:Antigens, CD34 / Q000032:analysis / Q000378:metabolism; D001854:Bone Marrow Cells / Q000737:chemistry* / Q000378:metabolism; D016026:Bone Marrow Transplantation; D001921:Brain / Q000098:blood supply / Q000378:metabolism; D001923:Brain Chemistry; D003287:Contrast Media / Q000737:chemistry; D003911:Dextrans; D052203:Ferrosoferric Oxide / Q000737:chemistry; D006801:Humans; D008252:Macaca fascicularis; D008279:Magnetic Resonance Imaging / Q000379:methods*; D058185:Magnetite Nanoparticles; D045164:Mesenchymal Stem Cell Transplantation; D059630:Mesenchymal Stem Cells / Q000737:chemistry* / Q000378:metabolism; D019408:Platelet Endothelial Cell Adhesion Molecule-1 / Q000032:analysis / Q000378:metabolism; D013194:Staining and Labeling / Q000379:methods*; D040301:Vascular Endothelial Growth Factor Receptor-2 / Q000032:analysis / Q000378:metabolism
D018952:Antigens, CD34; D003287:Contrast Media; D003911:Dextrans; D058185:Magnetite Nanoparticles; D019408:Platelet Endothelial Cell Adhesion Molecule-1; D040301:Vascular Endothelial Growth Factor Receptor-2; C065507:ferumoxides; D052203:Ferrosoferric Oxide