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Endogenous neural stem cell magnetic resonance noninvasive tracing technology system and establishment method

A technology of neural stem cells and tracing technology, which is applied in the field of magnetic resonance noninvasive tracing technology system and establishment of endogenous neural stem cells in the brain, and can solve the problems that cannot be used to analyze the metabolism of living tissue

Pending Publication Date: 2021-06-08
AFFILIATED HUSN HOSPITAL OF FUDAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Proton nuclear magnetic resonance spectroscopy (H-NMR) has been widely used in vitro to detect low levels of known metabolites and unknown compounds in body fluids and tissues; H-NMR can detect neuron-specific metabolites such as NAA and glial cells Unique products such as Cho; these complexes have been used as reliable biomarkers of the corresponding cell types in isolated tissues; however, practice has shown that H-NMR cannot be used to analyze the metabolism of substances in living tissues

Method used

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  • Endogenous neural stem cell magnetic resonance noninvasive tracing technology system and establishment method
  • Endogenous neural stem cell magnetic resonance noninvasive tracing technology system and establishment method
  • Endogenous neural stem cell magnetic resonance noninvasive tracing technology system and establishment method

Examples

Experimental program
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Effect test

Embodiment 1

[0019] Example 1 Isolation, culture and identification of rat embryo NSCs

[0020] (1) Primary culture of neural stem cells:

[0021] 14-16 day pregnant mice were selected and sacrificed after abdominal anesthesia with 0.32ml / 100g body weight with 10% chloral hydrate. Routine disinfection, laparotomy, fetal mice were taken out, and placed in 4°C PBS solution. After the fetal brain was taken out, the skull and meninges were removed, and the cortex and hippocampus were carefully separated under a dissecting microscope. Sharply cut hippocampus and cortex tissue to 1 mm by mechanical digestion 3 size, transfer to a 10ml cell centrifuge tube, add PBS to 3ml, carefully pipette and let stand for 2 minutes, suck out and collect the supernatant, add the same volume of PBS solution to the centrifuge tube containing the precipitated tissue, pipette again, and so on repeatedly, The tissue volume is gradually reduced until a single cell suspension is formed. Centrifuge at 800 rpm for 5 ...

Embodiment 2

[0037] Example 2 Isolation, cultivation and identification of NSCs in adult brain:

[0038] After debridement of patients with open brain injury, the exposed brain tissue was collected and placed in PBS solution, and washed with PBS repeatedly to remove blood clots and dirt. The pipette was blown to a single-cell suspension, and the cell concentration was adjusted to 5×10 with Neurobasal medium (Gibco) containing 10% FBS, 1% L-glutamine, 1% B27 and 20ng / ml EGF and bFGF. 5 / ml at 37°C, 5% CO 2 Suspension culture was carried out in a cell culture incubator, and fresh culture medium was replaced after 48 hours. Replace half of the culture medium every 3-4 days, and subculture once every 7-10 days. The identification method is the same as above.

Embodiment 3

[0039] Example 3 Stereotaxic transplantation of intracranial NSCs in rats:

[0040] Select the neural stem cell spheres cultured in vitro, trypsinize into a single cell suspension, and transplant 5ul (1X10 5 NSCs) in the right cerebral cortex of rats, the specific method: after the rats were anesthetized by intraperitoneal injection of 10% chloral hydrate (0.32ml / 100g), they were fixed on the KOPF stereotaxic apparatus in a prone position, and the scalp was incised in the middle, and right A 2mmX2mm bone window was formed 2mm laterally and 2mm posteriorly, about 2.5mm vertically from the dura mater, and 5 μL containing about 1×10 5 Rat NSCs suspension, and keep the needle at the injection site for 5 minutes to prevent leakage, and then gradually pull out the injection needle within 3 minutes. After the transplantation, the scalp was sutured, 2000 units of gentamicin were injected intraperitoneally for antibacterial treatment, and the rats were returned to their cages for feed...

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Abstract

The invention belongs to the technical field of biomedicine, and particularly relates to an intracerebral endogenous neural stem cell magnetic resonance noninvasive tracing technology system and an establishment method. The spectrum characteristics of neural stem cells are analyzed in vitro to obtain the spectrum characteristics, the peak value of the spectrum characteristics can be used as a biological marker of the neural stem cells in vivo and vitro, after animal living body level verification, endogenous neural stem cells in a human brain are detected at the living body level, and the result shows that a small amount of neural stem cells exist in a hippocampus area, and the number of the neural stem cells is reduced along with age increase. According to the tracing technology system, the distribution and biological behaviors of endogenous neural stem cells in the brain can be traced in the living body level by using MRS (magnetic resonance spectroscopy), and the technology system provides a visual and noninvasive technical method for clinical neuroscience and clinical evaluation of nerve regeneration, regeneration indexes and the like in the brain, and has important clinical value and significance.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and relates to an imaging tracing technology system of endogenous neural stem cells in the brain, in particular to a magnetic resonance non-invasive tracing technology system and establishment method of endogenous neural stem cells in the brain. The technical system can use MRS to trace the biological behavior of endogenous neural stem cells in the brain at the living level. Background technique [0002] Studies have revealed the ability in the adult mammalian brain to generate new neurons from neural stem cells (NSCs) located in the lateral subventricular zone and hippocampus, which have the ability to self-renew and generate daughter cells. Studies have shown that NSCs can generate neurons, astrocytes and oligodendrocytes in vivo and in vitro, which provides the possibility of using NSCs to repair damaged or missing nerve tissue due to neurodegenerative diseases and trauma. At present, the...

Claims

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Application Information

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IPC IPC(8): A61B5/055A61D1/00C12N5/0797G01N27/62
CPCA61B5/055A61D1/00C12N5/0623G01N27/62C12N2509/00C12N2500/32C12N2501/11C12N2501/115C12N2500/02
Inventor 朱剑虹汤海亮李荣刚朱侗明吴惺胡锦
Owner AFFILIATED HUSN HOSPITAL OF FUDAN UNIV
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