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A kind of exosome with diagnosis and treatment function and preparation method thereof

A technology of exosomes and functions, applied in the field of nanomedicine, can solve the problems of insufficient timeliness and accuracy in tumor diagnosis and condition evaluation, and achieve the effects of improving solubility, short preparation time, and easy operation

Active Publication Date: 2021-12-21
绿钥生物科技(广州)有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Traditional tumor treatment can only inhibit tumor growth through drugs, but the diagnosis and condition evaluation inside the tumor are not timely and accurate enough

Method used

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  • A kind of exosome with diagnosis and treatment function and preparation method thereof
  • A kind of exosome with diagnosis and treatment function and preparation method thereof
  • A kind of exosome with diagnosis and treatment function and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Add 80 μg of exosomes to 10 mM PBS solution, mix well, add to a 96-well plate, put it into an electroporation instrument, and operate according to the following parameters: voltage 250 V, pulse width 100 μs, interval 1000 μs, discharge times 6 times. After electroporation, put the suspension in a cell culture incubator and incubate for 30 minutes, then ultracentrifuge twice at 100,000g for 70 minutes each time, remove the supernatant, and resuspend the pellet with 200 μL PBS to obtain exosomes after electroporation. Its particle size is ~150nm, such as image 3 shown. Zeta potential is ~-8mV, such as Figure 4 shown. Store at 4°C in the dark for subsequent testing and experiments.

Embodiment 2

[0045] Add 80 μg of exosomes and 80 μL of curcumin solution (concentration: 2 mg / mL) into 10 mM PBS solution, mix well, add to a 96-well plate, put it into an electroporation apparatus, and operate according to the following parameters: voltage 250 V, pulse width 100 μs , the interval is 1000μs, and the number of discharges is 6 times. After electroporation, put the suspension in a cell culture incubator and incubate for 30 minutes, then ultracentrifuge twice at 100,000 g for 70 minutes each time, remove the supernatant, and resuspend the pellet with 200 μL PBS to obtain curcumin-loaded exosomes. Its encapsulation efficiency is ~86.64%, such as figure 2 Shown; its particle size is ~ 170nm, such as image 3 Shown; Zeta potential is ~-17mV, such as Figure 4 shown; the prepared exosomes loaded with curcumin have drug sustained release properties, such as Image 6 shown. Store at 4°C in the dark for subsequent testing and experiments.

Embodiment 3

[0047] Add 80 μg of exosomes and 40 μL of indocyanine green solution (concentration: 2 mg / mL) into 10 mM PBS solution, mix well, add to a 96-well plate, put it into an electroporation apparatus, and operate according to the following parameters: voltage 250 V, pulse The width is 100μs, the interval is 1000μs, and the number of discharges is 6 times. After electroporation, place the suspension in a cell culture incubator and incubate for 30 minutes, then ultracentrifuge twice at 100,000 g for 70 minutes each time, remove the supernatant, and resuspend the pellet with 200 μL PBS to obtain indocyanine green-loaded exocytosis. body, its encapsulation efficiency is ~97.79%, such as figure 2 Shown; the particle size is ~180nm, such as image 3 Shown; Zeta potential is ~-6mV, such as Figure 4 shown. The prepared exosomes loaded with indocyanine green have fluorescence imaging, photothermal effect and drug sustained release properties, such as figure 1 , 5, 6. Store at 4°C in t...

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Abstract

The invention discloses an exosome with diagnostic and therapeutic functions and a preparation method thereof, belonging to the technical field of nanomedicine. The method includes: 1) exosome extraction and purification: extract exosomes secreted by the cells from the culture medium of C57 mouse mesenchymal stem cells, resuspend the obtained exosomes in 10mM PBS solution, and store them at -80°C for future use ; 2) Sample preparation: Mix the prepared curcumin or indocyanine green solution with the exosomes in step 1) in 10mM PBS to obtain a sample; 3) Electroporation: electroporate the sample in step 2) to obtain The sample after electroporation; 4) Incubation: Place the electroporated sample in step 3) in a cell culture incubator and incubate for 30 minutes in the dark to obtain the incubated liquid; 5) Purification: centrifuge the incubated liquid in step 4) for 60 After ‑100min, the precipitate was collected to obtain exosomes with diagnostic and therapeutic functions. The invention has simple operation and high repetition rate.

Description

technical field [0001] The invention belongs to the technical field of nanomedicine, and in particular relates to an exosome with diagnostic and therapeutic functions and a preparation method thereof. Background technique [0002] Chemotherapy is still one of the main ways of cancer treatment. However, most chemotherapeutic drugs have defects such as poor water solubility, rapid metabolism, poor biocompatibility, unsatisfactory distribution in the body, and low cell penetration ability, which severely limit the efficacy and utilization of chemotherapeutic drugs. Therefore, it is a current research hotspot to construct a new type of drug carrier to improve drug solubility and stability, improve tissue penetration, increase drug concentration at tumor sites, and avoid being captured by immune cells. [0003] The unique advantages of exosomes in drug delivery make them one of the research hotspots. Exosomes are nanovesicles derived from mammalian cells, with a diameter of abo...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K9/127A61K47/46A61K31/12A61K41/00A61K49/00A61P35/00
CPCA61K41/0052A61K41/0057A61K47/46A61K49/0034A61K49/005A61P35/00A61K9/1271A61K31/12A61K2300/00
Inventor 谢茂彬李广蒙王金恒
Owner 绿钥生物科技(广州)有限公司
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