A kind of exosome sustained-release polypeptide hydrogel and preparation method and use thereof
A hydrogel and exosome technology, applied in the field of exosome slow-release polypeptide hydrogel, can solve problems such as short half-life, achieve slow release, prolong half-life, and have good clinical application prospects
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Embodiment 1
[0033] Example 1 Preparation of ExOSomes of the MSCS source of the present invention
[0034] The preparation method is as follows:
[0035] 1, MSCS primary culture: mouse is broken, 70% alcohol soaked for 10 minutes, separating bilateral tibia, rinsing bone marrow with serum-free medium, collecting rinsing liquid, centrifuging, and using an adhesive method to obtain MSCs.
[0036] 2. Culture Collection: DMEM medium (US GBICO) developing MSCs (US GBICO) was cultured in DMEM medium (US GBICO), US GBICO, US SBI, and regular collection.
[0037] 3, centrifugal extraction: Extraction of EXOS, 2000 g × 10 minutes by multiple centrifugation methods, and remove cell fragments.
[0038] 4, purification: The last EXOS was cleaned once with PBS, and then centrifuged 10,000 degrees for 60 minutes again.
Embodiment 2
[0039] Example 2 Preparation of a polypeptide hydrogel of the present invention
[0040] According to the treatment of the present invention, the hydrogel sustained EXOS is used, and the sustained release system is determined according to the pre-test results (therapeutic effect) and the experimental animal (C57 mouse) and the application site (mouse kidney film) as follows:
[0041] Element Concentration (mg / ml) Volume (μL) EXOSMES (EXOS) 8 10 Self-assembled short peptide KMP2 10 13 Sterile water 10 3
2 total 25
[0042] (1) Self-assembled short peptide KMP2 (KLDLPVGLIGKLDL) was synthesized from Shanghai Potati Biotechnology, and dried from dry powder in sterile water, and was prepared from 10 mg / ml of reservoir, and 13 μL was added to the 1.5 mL of EP tube;
[0043] (2) Take 10 μl of concentration of 8 mg / ml EXOS to EXOS to add EP tube, mix well.
[0044] (3) 2 μl of sterile water, blow mixing mixed.
[0045] Example 2 Preparation...
Embodiment 3
[0048] Example 3 Preparation of Polypeptide Watergels of Cogens
[0049] (1) Self-assembled short peptide KMP2 (KLDLPVGLIGKLDL) is purified by Shanghai Potati Biotechnology, and 300 ug self-contained peptide is dissolved in 20 μl of water into a self-assembled peptide aqueous solution; 100 ug of exosomes are dissolved in 15 μL of water formation into an outer body Aqueous solution; add in the EP tube;
[0050](2) The self-assembled peptide solution obtained from step (1), an extractive solution, add 5 μl of water, a total of 40 μl, mix average, and a polypeptide hydrogel.
[0051] The beneficial effects of the present invention are proved by specific pharmacodynamics experiments:
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