Preparation of an exosome-doxorubicin nano-targeted drug derived from bone marrow mesenchymal stem cells and its anti-osteosarcoma effect in vitro

A bone marrow mesenchymal, exosome technology, applied in the direction of bone/connective tissue cells, animal cells, drug combinations, etc., to achieve the effects of low immunogenicity, strong anti-tumor efficacy, and strong expansion ability

CN110124058BActive Publication Date: 2021-11-26THE FIRST AFFILIATED HOSPITAL OF FUJIAN MEDICAL UNIV
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Patents(China)
Current Assignee / Owner
Publication Date
2021-11-26

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Abstract

The invention discloses the preparation of an exosome-doxorubicin nano-targeted drug derived from bone marrow mesenchymal stem cells and the research on anti-osteosarcoma in vitro. It includes the following steps: (1) Culture BM-MSCs in serum-free medium, and extract exosomes with an exosome extraction kit. (2) Mix exosomes and doxorubicin hydrochloride in a certain proportion, add triethylamine for desalting, and then place the mixture in a dialysis bag at 4°C overnight to prepare exosome-doxorubicin drug. (3) Place the prepared drug in a microplate reader, measure its absorption peak at 490 nm, and calculate the drug loading capacity. (4) The prepared drug was used in the curative effect research of anti-osteosarcoma MG63 cells in vitro. The MSCs selected by this method have the advantages of less toxic and side effects in vivo, low immunogenicity, tumor targeting, and large-scale expansion in vitro. It can effectively enhance the killing effect of doxorubicin on osteosarcoma cells.
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Description

technical field

[0001] The invention relates to a preparation method of exosomes derived from bone marrow mesenchymal stem cells-doxorubicin nano-targeted drug and research on anti-osteosarcoma. Background technique

[0002] Drug carriers refer to natural or synthetic polymer materials, which are chemically bonded, physically adsorbed or packaged with drug molecules in different forms to form a drug control system. Due to the advantages of small particles, high stability and targeting, nano drug carriers have been paid more and more attention by researchers. At present, many nanomaterials are used as nano-drug carriers, such as gold nanoparticles, carbon nanotubes, nanomicelles, liposomes, and polymer nanoparticles. Among these nanocarriers, liposome-based nanocarriers have good biocompatibility and degradability, the phospholipid membrane is easy to break through the plasma membrane, it is easy to separate the drug from the surrounding environment, and it can be combined w...

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Embodiment Construction

[0019] In order to make the technical problems, technical solutions and beneficial effects to be solved by the present invention clearer, the present invention will be further described in detail below in conjunction with the embodiments. It should be understood that the specific embodiments described here are only used to explain the present invention, not to limit the present invention.

[0020] The embodiment of the present invention is achieved in this way. The preparation of exosomes derived from bone marrow mesenchymal stem cells-doxorubicin nano-targeted drug and the research on anti-osteosarcoma in vitro include the following steps:

[0021] (1) Cultivation of bone marrow mesenchymal stem cells: When the mesenchymal stem cells grow to a confluence of 70%-80%, replace the serum-free medium and culture for 48 hours, and then collect the medium for exosome extraction.

[0022] (2) Extraction of exosomes: Centrifuge the collected culture at 3000g for 10 minutes at 4°C to r...