Nano probe transfected adipose-derived cell and application thereof in preparing MRI (magnatic resonance imaging) dynamic tracer
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A fat source and tracer technology, applied in the field of cell biology, can solve unseen problems
Inactive Publication Date: 2014-01-29
SHANGHAI NINTH PEOPLES HOSPITAL AFFILIATED TO SHANGHAI JIAO TONG UNIV SCHOOL OF MEDICINE
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The nanoprobe can effectively label and trace bone marrow stromal stem cells, neural stem cells, embryonic stem cells, etc. in vivo or in vitro, but there is no relevant report on the repair of unstable plaques of atherosclerosis by using nanoprobes to trace EPCs
[0006] In the previous research work, the inventors have mastered and established methods for the isolation, culture and expansion of C57 and GFP mouse adipose-derived EPCs and human umbilical cord-derived EPCs, and induced their differentiation into endothelial cells in vitro, successfully developed an independent Nano-pro
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Embodiment 1
[0043] (1) Isolation, cultivation, expansion and identification of adipose tissue-derived cells
[0044] Newborn SD mice aged 3-7 days were taken, killed by dislocation, put in 75% alcohol, and soaked for 5-10 minutes; under sterile conditions, bilateral inguinal fat was taken; cut into pieces, digested and filtered, sorted by flow cytometry, Immunomagnetic bead sorting and differential adherence digestion method to purify and obtain CD34+ cells with morphological characteristics of endothelial progenitor / cells, and perform immunofluorescence identification and flow cytometry identification (CD34 / CD31 / KDR / CD90, etc.) Phenotype expression and cell purity, and induce differentiation to endothelial cells;
[0045] (2) Preparation of dendrimer functionalized nano-molecular probes and functional detection.
[0046] Preparation of iron oxide nanoparticles (Iron Oxide nanoparticles) with a diameter of 5-50nm, polyglutamic acid (PGA) and poly-L-lysine (PLL) surface shell crosslinking...
Embodiment 2
[0049] Example 2 MRI tracer test of nano-molecular probe-labeled adipose-derived cells repairing vascular damage in different periods:
[0050] (1) In vivo tracking of adipose-derived cells repairs atherosclerotic unstable plaques in Apo E mice
[0051] The adipose-derived cells of GFP mice labeled with different concentrations of nanoprobes were injected into ApoE gene-deficient 9-month-old mice through the tail vein, and the homologous C57 mice were used as the control group. MRI scanning was performed on 30d, 60d, and 90d to observe the development of the above-mentioned particles in the unstable plaque of atherosclerosis, and measure the R2 and R2* values of the unstable plaque; 7d, 14d, 30d, 60d, and 90d after operation Histological observations (HE, immunofluorescent staining, Prussian blue staining, etc.) were performed after the samples were collected, and their ability to repair unstable plaques of atherosclerosis was evaluated; at the same time, the transplanted ce...
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Abstract
The invention belongs to the field of cytobiology, and relates to a nano probe transfected adipose-derived cell and application thereof in preparing an MRI (magnatic resonance imaging) dynamic tracer. The adipose-derived progenitor/stem cell is separated and screened from adipose tissues, the phenotype of the adipose-derived progenitor/stem cell is identified by stream mode, immune fluorescence, molecular biology and other methods, and the inward epidermic cell differentiation is induced. After the nanoparticle labeled adipose-derived endodermis progenitor/stem cell is prepared into an MRI dynamic tracer, the in-vivo transplantation vascular injury restoration test, which is used for evaluating vascular endodermis conditions by transmission electron microscope, immunofluorescence, real-time PCR (polymerase chain reaction) and other methods according to the in-vivo survival, migration, outcome and restoration vascular injury conditions of the nuclear magnetic resonance dynamic tracing adipose-derived endothelium progenitor/stem cell at different time points, proves that the nano probe transfected adipose-derived cell can illuminate the mechanism of the adipose-derived endodermis progenitor/stem cell in vascular injury restoration. The nano probe transfected adipose-derived cell can provide new practice data for vascular injury treatment as well as new seed cell sources and experimental reference.
Description
technical field [0001] The invention belongs to the field of cell biology, and relates to adipose-derived cells transfected with nanoprobes and its application, in particular to a method for separating and obtaining adipose-derived cells from adipose tissue, transfecting them with nanoprobes and preparing MRI dynamics. Use in tracer. Background technique [0002] It is known that the heart and cerebrovascular injury caused by atherosclerosis (AS) seriously affects and threatens human health; in an aging society, heart and cerebrovascular diseases have become the number one cause of human death. Therefore, the prevention and treatment of AS and vascular injury has always been the focus and difficulty of research. The traditional view is that lipid deposits in the subintima of arteries and atherosclerotic plaques progress, resulting in high stenosis of vascular lumens, tissue ischemia and hypoxia, and ultimately acute cardiovascular and cerebrovascular events. However, in re...
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IPC IPC(8): C12N5/077A61K49/14A61K49/12A61K49/08
Inventor 陆信武秦金保叶开创李祥祥蒋米尔
Owner SHANGHAI NINTH PEOPLES HOSPITAL AFFILIATED TO SHANGHAI JIAO TONG UNIV SCHOOL OF MEDICINE
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