Super-paramagnetic nanometer vector carrying retinal pigment epithelium derived factor (PEDF)
A retinal pigment and epithelial cell technology, applied in the field of superparamagnetic nanocarriers, can solve the problems of little understanding of potential toxicity, reduction, and low relaxation
Inactive Publication Date: 2011-04-13
AFFILIATED HUSN HOSPITAL OF FUDAN UNIV
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Problems solved by technology
Although gadolinium chelating agents can be used for MR molecular or cellular imaging, the relaxation of such contrast agents is low, and the internalization of cells will be further reduced. Moreover, gadolinium chelating agents do not have biocompatibility. Toxicity is poorly understood
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Embodiment 1
[0028] 1. Preparation of surface functionalized superparamagnetic iron nanoparticles (USPIO)
[0029] Synthesis of Nano Fe by Chemical Co-precipitation 3 o 4 Magnetic ions, surface coated with dextran, modified with carboxylated functional groups, and using transmission electron microscopy to display nano-magnetic particles, see figure 1 .
[0030] 2. Synthetic human AAV-PEDF cross-linked with USPIO to make a gene carrier (PEDF-USPIO)
[0031] 1) Preparation of PEDF-USPIO carrier:
[0032] The PEDF cDNA amplified by PCR was digested with BamHI and NcoI, and connected into the eukaryotic expression vector AAV-GFP to construct the AAV-PEDF expression vector. After double-enzyme digestion and sequencing (attached figure 2 ), take 1 mg of magnetic microspheres and 20 μg of AAV-PEDF, disperse them in 0.6 ml of cross-linking buffer, shake the reaction at 50°C for 3 hours, wash the magnetic microspheres with citric acid buffer at room temperature for 3 times, and wash them with...
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Abstract
The invention belongs to the technical field of biology and relates to a super-paramagnetic nanometer vector (USPIO) carrying retinal pigment epithelium derived factor (PEDF). In the invention, super-paramagnetic nanometer iron particles are prepared through a nanometer technology and the surfaces of the particles are functionally modified; the retinal pigment epithelium derived factor is constructed on an adenovirus related expression vector AAV-GFP containing green fluorescent protein report gene so as to crosslink USPIO with the AAV-GFP; and glioma cells are induced in vitro and devoured. By utilizing the magnetic marker function of the vector of the invention, the influences of the PEDF on the growth, the differentiation and the invasiveness of the brain glioma are noninvasively monitored in vivo, thereby breaking new incorporating and monitoring ways for the clinical application of the glioma gene treatment, and providing innovative operation means and gathering exploratory experience for the research of the clinical anti-tumor treatment.
Description
technical field [0001] The invention belongs to the field of biological technology, and in particular relates to a superparamagnetic nanocarrier (PEDF-USPIO) carrying retinal pigment epithelial cell-derived factor (PEDF). Background technique [0002] Studies have shown that glioma is derived from glial cells in the brain and is the most common malignant brain tumor, accounting for about 50% of intracranial tumors. Its malignant phenotype is characterized by rapid proliferation, local invasion and angiogenesis . Although a lot of research work on the treatment of glioma is going on, there has been no obvious breakthrough in the mechanism of its occurrence, development and deterioration and related treatment methods. Clinical practice shows that even after surgery, postoperative radiation therapy, chemotherapy, and even immature biological therapy, recurrence and progression are still inevitable for most patients. hot spots of our attention. [0003] Retinal pigment epithe...
Claims
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IPC IPC(8): C12N15/861C12Q1/02
Inventor 张弢
Owner AFFILIATED HUSN HOSPITAL OF FUDAN UNIV