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Neural stem cells traced by fluorescence and SPECT/CT double-image functional microspheres and application

A neural stem cell and dual imaging technology, applied in neural stem cells and application fields, can solve the problem of undiscovered molecular probes, and achieve the effects of good biodegradability and biocompatibility, low cytotoxicity and uniform particle size distribution

Active Publication Date: 2017-11-24
SOUTHEAST UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

So far, no molecular probes with optical properties based on iridium compounds have been found to track NSCs and perform imaging observations.

Method used

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  • Neural stem cells traced by fluorescence and SPECT/CT double-image functional microspheres and application
  • Neural stem cells traced by fluorescence and SPECT/CT double-image functional microspheres and application
  • Neural stem cells traced by fluorescence and SPECT/CT double-image functional microspheres and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] The preparation method of fluorescent and SPECT / CT dual-image functional microspheres includes the following steps:

[0044] (1) Weigh 2 mg (Bis(2-methyldibenzo[f,h]quinoxaline)(acetylacetonate)iridium(III))(Ir(MDQ) 2 acac) and 25 mg of polyglycolide-polyethylene glycol-copolymer (PLGA-PEG) were mixed, dissolved in 1 mL of chloroform to obtain an oil phase, and 120 μL of I was prepared 125 Na was used as the internal water phase.

[0045] (2) ultrasonically mixing the oil phase obtained in step (1) and the inner water phase for 5 times (2s, 100W, BILON92-II) to obtain a colostrum.

[0046] (3) Add 4 mL of PVA (polyvinyl alcohol, poly vinyl alcohol) aqueous solution (5 wt %) to the product of step (2), and ultrasonically mix 5 times (2s, 100W) to form a double emulsion.

[0047] (4) The product of step (3) was diluted into an aqueous solution of PAA (polyacrylic acid, poly acrylic acid) (5 wt %, 40 mL), stirred overnight at room temperature, and evaporated in the dark....

Embodiment 2

[0052] (1) NSCs are subcultured normally and cultured in suspension in non-adherent culture flasks;

[0053] (2) Mix the microsphere solution obtained in Example 1 with the cell culture medium to prepare a microsphere culture solution (0.04-0.8 mg / mL) and place it in an incubator (37°C, 5% CO 2 ) for 24h.

[0054] (3) After 24 hours, the neural stem cells proliferate normally, add the same volume of the iridium compound-containing microsphere culture medium (0.04-0.8 mg / mL) prepared in step (2) to make the final concentration of the microspheres after the cells are mixed with the microspheres 0.02-0.4 mg / mL, mix the microspheres with the cells and place them in an incubator (37°C, 5% CO 2 ), continue to cultivate for 1-3 days, preferably 3 days.

[0055] (4) After 1-3 days, centrifuge at 1000rpm for 5min, remove the culture medium, wash twice with PBS, inoculate the cells into 24-well glass slides pre-incubated with laminin (laminin) for 12h and continue to incubate for 12h,...

Embodiment 3

[0064] (1) 0.2 mg of Ir compound was weighed, mixed with 25 mg of polyglycolide, polylactic acid-polyethylene glycol copolymer, and polyglycolide-polyethylene glycol copolymer, respectively, and then dissolved in 1 mL of chloroform, respectively. To get the oil phase, prepare 10 μL of I 125 Na as internal water phase;

[0065] (2) ultrasonically mixing the oil phase and the inner water phase obtained in step (1) for 5 times (2s, 100W, BILON92-II) to obtain a colostrum;

[0066] (3) adding 4 mL of PVA aqueous solution (5wt%) to the product of step (2), and ultrasonically mixing 5 times (2s, 100W) to form a double emulsion;

[0067] (4) The product of step (3) was diluted into PAA aqueous solution (5wt%, 40mL), stirred overnight at room temperature, and volatilized in the dark;

[0068] (5) Use a 50mL high-speed centrifuge tube to collect step (4) The solution containing microspheres, first wash with absolute ethanol, centrifuge at high speed (14500rpm, 20min), discard the sup...

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Abstract

The invention relates to neural stem cells traced by fluorescence and SPECT / CT double-image functional microspheres. The neural stem cells endocytose the fluorescence and SPECT / CT double-image functional microspheres. The fluorescence and SPECT / CT double-image functional microspheres comprise the following components, by mass, of 70%-99.9% of polymers, 0.05%-20% of cyclic metal iridium compounds and 0.05%-10% of I125 agents, wherein the polymers are degradable. The invention further discloses an application of the neural stem cells in preparation of tracers and / or therapeutic agents for a spiral neuronal injury or a neurodegenerative disease. According to the prepared neural stem cells traced by the fluorescence and SPECT / CT double-image functional microspheres, a noninvasive, dynamic, visual, simple and convenient method is provided for tracing tissue engineering seed cells implanted into a living body or a human body, and an observing and repairing technology aiming at the neurodegenerative disease is provided.

Description

technical field [0001] The invention relates to the technical field of molecular probes, in particular to neural stem cells traced by fluorescent and SPECT / CT double image function microspheres and its application. Background technique [0002] At present, neurodegenerative disease is a kind of chronic progressive disease. Although the lesion sites and causes of different types of neurodegenerative diseases are different, the degeneration of neuron cells in specific parts of the brain is a common feature of these diseases. Neurodegenerative diseases such as Alzheimer's disease and Parkinson's disease seriously affect human work and life. Neuronal cells in the brain generally do not regenerate, and neurodegenerative diseases worsen over time, eventually leading to dysfunction. The pathogenesis of neurodegenerative diseases has not been confirmed, but a large number of studies have pointed out that neuron cell damage and loss are the key factors leading to its pathogenesis. ...

Claims

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Application Information

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IPC IPC(8): C12N5/0797A61K35/30A61K49/00A61K49/04A61K51/06A61K51/12A61P25/00A61P27/16A61K101/02
CPCA61K35/30A61K49/0002A61K49/0019A61K49/0054A61K49/0056A61K49/0091A61K49/04A61K49/048A61K51/06A61K51/1251C12N5/0623
Inventor 李丹唐明亮邱裕友顾炜周宇杨张琦
Owner SOUTHEAST UNIV
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