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Method for preparing mesenchymal stem cell exosome and application thereof

A technology of stem cells and exosomes, which is applied in the field of stem cells and regenerative medicine, can solve the problems of low concentration, low extraction efficiency, and complicated ultracentrifugation operation, and achieve the effects of increasing concentration, saving separation time and cost, and reducing volume

Inactive Publication Date: 2018-12-28
GUANGDONG VITALIFE BIOTECHNOLOGY CO LTD
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Problems solved by technology

[0005] The purpose of the present invention is to provide a method for preparing mesenchymal stem cell exosomes, aiming at technical problems such as low concentration of mesenchymal stem cell exosomes, complicated ultracentrifugation operation, and low extraction efficiency. High secretion concentration, easy operation of ultracentrifugation, high extraction efficiency

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  • Method for preparing mesenchymal stem cell exosome and application thereof
  • Method for preparing mesenchymal stem cell exosome and application thereof
  • Method for preparing mesenchymal stem cell exosome and application thereof

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Embodiment Construction

[0031] The technical solutions of the present invention will be described in further detail below in conjunction with specific examples, but the present invention is not limited to the following examples.

[0032] Unless otherwise specified, the reagents used in the present invention are all existing reagents, which can be obtained through commercial channels.

[0033] Follow the steps below to prepare injections:

[0034] 1. The hospital collects the umbilical cord tissue of the full-term caesarean section fetus, signs the customer's informed consent before the collection, and transports the tissue to the laboratory in a refrigerated and sterile environment at 4°C. During the transportation, the tissue protection solution is used to protect the biological activity of the umbilical cord tissue, and the tissue protection solution Prepared by adding 25 μg / ml gentamicin sulfate and 5 μg / ml amphotericin B to normal saline to ensure no bacterial and fungal contamination during tran...

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Abstract

The invention provides a method for preparing a mesenchymal stem cell exosome. The method comprises the steps that human umbilical cord tissue surface blood is cleaned to remove epidermal and vasculartissues, Wharton jelly is taken out for cleaning, then the tissues are cut into pieces, and inoculation is performed for culture; subculturing is performed; second generation of umbilical mesenchymalstem cells are inoculated and cultured; a cell stimulator factor for promoting secretion of the exosome is added; when the cells grow to the fusion degrees 90%, first cell supernate is sucked out forseparating the exosome; the first cell supernate is centrifuged to remove cell debris, and second cell supernate is collected; the first time of ultrafiltration and concentration is conducted on thesecond cell supernate, excessive moisture is removed, and a first concentrated solution is obtained; the first concentrated solution is subjected to the second time of ultrafiltration and concentration, excessive moisture is removed, and a second concentrated solution is obtained; the first time of ultracentrifugation is conducted on the second concentrated solution, and the supernate is taken andsubjected to the second time of ultracentrifugation, the supernate is removed, and precipitation is performed to obtain the exosome for stem cell secretion; normal saline is added for cleaning and centrifugation, the supernate is removed, and the exosome precipitate is obtained.

Description

technical field [0001] The invention belongs to the field of stem cells and regenerative medicine, and in particular relates to a method for preparing mesenchymal stem cell exosomes and an application thereof. Background technique [0002] Exosome (Exosome) is a kind of membranous vesicle released from the intracellular vesicle structure to the outside of the cell, with a diameter of about 30-150nm. Cells will secrete exosomes under normal physiological conditions, through the protein in the vesicle RNA and RNA play an important role in intercellular signal transduction and intercellular regulation, and the mechanism of this signal transduction and mutual regulation is complex, especially between tumor cells, whether the exosomes secreted by tumor cells can affect the Immunosuppression, tumor cell migration, proliferation, and infiltration have promoting effects, etc., which are not yet fully understood. [0003] A bottleneck problem that needs to be solved urgently in exos...

Claims

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Application Information

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IPC IPC(8): C12N5/0775
CPCC12N5/0662C12N5/0665C12N2500/38C12N2501/115C12N2501/18C12N2501/24
Inventor 朱建斌江嘉豪何美弟陈炽峰王进辉于莉
Owner GUANGDONG VITALIFE BIOTECHNOLOGY CO LTD
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