A hypoxia-responsive co-assembly system based on extracellular vesicles and preparation method thereof

A co-assembly and system technology, applied in the biological field, can solve problems such as special equipment complexity space, body death, body damage, etc.

Active Publication Date: 2022-07-22
NANKAI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, excessive hypoxic signals can lead to body damage and even death
In view of the deepening understanding of hypoxia on diseases, researchers are paying more and more attention to the clinical application of tracer tissue hypoxia, such as magnetic resonance imaging (MRI) and positron emission tomography (PET), etc. , but due to the complexity of special equipment and its limited space, the use of these large instruments in scientific research still has certain inconvenience

Method used

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  • A hypoxia-responsive co-assembly system based on extracellular vesicles and preparation method thereof
  • A hypoxia-responsive co-assembly system based on extracellular vesicles and preparation method thereof
  • A hypoxia-responsive co-assembly system based on extracellular vesicles and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0075] This example is used to illustrate a method for extracting extracellular vesicles derived from umbilical cord-derived mesenchymal stem cells.

[0076] The processing steps are as follows: place fetal bovine serum in an ultracentrifuge tube, centrifuge at 100,000g at 4°C for 2 hours, take the supernatant in an ultra-clean bench, filter with a 0.22 μm needle filter, and store it in a -80°C refrigerator for later use.

[0077] For cell biology experiments such as cell subculture, cryopreservation and recovery, please refer to "Animal Cell Culture (Sixth Edition)".

[0078] Conditioned medium for collecting human umbilical cord-derived mesenchymal stem cells (hP-MSCs) containing extracellular vesicles: when cultured at 75 cm 2 The hP-MSCs in the cell culture flask were in the logarithmic growth phase, and when the cell confluence reached 80%, the medium was exhausted, washed twice with PBS, and 10 ml of prepared 10% extracellular vesicle-free solution was added to each flas...

Embodiment 2

[0085] This example is used to illustrate a method for preparing a co-assembly system based on extracellular vesicles (EVs) for hypoxia-responsive imaging.

[0086] (1) After mixing aluminum phthalocyanine Pc and calixarene C5A respectively, they were treated in a dark room at 37°C for 30 min.

[0087] (2) Add Pc / C5A to the extracted 100 μL EVs sample containing 200-300 μg protein, make up to 500 μL with PBS, at this time the final concentration of Pc / C5A is 10 μM / 20 μM, invert and mix, and warm at 37°C Incubate for 2h.

[0088] (3) The above mixture was transferred into an ultracentrifuge tube, filled with PBS, centrifuged at 100,000 g at 4° C. for 120 minutes, and the supernatant was removed to obtain green-stained extracellular vesicles.

[0089] (4) Resuspend the extracellular vesicles in 50 μl PBS, aliquot and store in a -80°C refrigerator for later use.

Embodiment 3

[0091] This example is used to illustrate a method for the identification of a hypoxia-responsive imaging co-assembly system based on extracellular vesicles (EVs).

[0092] (1) Identification of extracellular vesicle morphology using transmission electron microscopy

[0093] The extracellular vesicles extracted in Example 1 and the hypoxia response imaging co-assembly system of Example 2 were dropped on a 200-mesh sample copper grid, kept at room temperature for 2 min, and the excess liquid was blotted with filter paper; 20 mg / mL was added dropwise on the sample grid. The uranyl acetate solution was allowed to stand at room temperature for 1 min, the samples were negatively stained, the excess liquid was blotted with filter paper, and the sample net was air-dried; the prepared samples were observed under a transmission electron microscope, and photographs were collected. like figure 1 As shown, the shape and diameter of extracellular vesicles did not change, and the shape was...

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Abstract

The present disclosure relates to a co-assembly system for hypoxia response imaging based on extracellular vesicles, the co-assembly system comprising extracellular vesicles and Pc / C5A attached to the extracellular vesicles, the Pc / C5A contains calixarene QAC5A‑6C and aluminum phthalocyanine AlPcS 4 ; the calixarene QAC5A-6C (C5A) and aluminum phthalocyanine AlPcS 4 (Pc) A first incubation mixed and performed yields Pc / C5A; a second incubation mixed with extracellular vesicles results in a co-assembled system for hypoxia-responsive imaging.

Description

technical field [0001] The present disclosure relates to the field of biotechnology, in particular, to a co-assembly system for hypoxia response imaging based on extracellular vesicles, and a method for preparing a co-assembly system for hypoxia response imaging based on extracellular vesicles Methods and a self-assembled Pc / C5A based on co-assembly with extracellular vesicles for imaging of hypoxia response. Background technique [0002] Oxygen is an essential element in the energy metabolism of all living organisms, and many physiological processes are regulated by oxygen, which makes the body's perception and regulation of changes in oxygen concentration in the environment very precise. Hypoxia is a key cause of many diseases, such as tumors, anemia, cardiovascular and cerebrovascular diseases, etc. In fact, mild hypoxia mainly causes a compensatory response in the living body, and the hypoxia signaling pathway has a protective effect on oxidative damage and inflammation...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K49/00
CPCC09K11/06C09K11/025A61K49/0036A61K49/0052A61K49/005G01N21/6428C09K2211/1074C09K2211/186
Inventor 王悦冰郭东升程元秋岳宇昕王斓星
Owner NANKAI UNIV
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