Preparation method and storage method for placenta-derived mesenchymal stem cell exosome

A technology of mesenchymal stem cells and storage methods, applied in the field of preparation and storage of exosomes from placental mesenchymal stem cells, can solve the problems of time-consuming, vesicle damage, unstable recovery rate, etc., and achieve high activity and function effects

Inactive Publication Date: 2020-05-01
GUANGDONG VITALIFE BIOTECHNOLOGY CO LTD
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

However, the most commonly used method for the separation and extraction of exosomes from placental mesenchymal stem cells is ultracentrifugation, which uses low-speed and high-speed centrifugation alternately, but this method

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  • Preparation method and storage method for placenta-derived mesenchymal stem cell exosome
  • Preparation method and storage method for placenta-derived mesenchymal stem cell exosome
  • Preparation method and storage method for placenta-derived mesenchymal stem cell exosome

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Embodiment Construction

[0039]In order to facilitate the understanding of the present invention, the present invention will be described more fully below with reference to the associated drawings. Preferred embodiments of the invention are shown in the accompanying drawings. However, the present invention can be embodied in many different forms and is not limited to the embodiments described herein. On the contrary, these embodiments are provided to make the understanding of the disclosure of the present invention more thorough and comprehensive.

[0040] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the technical field of the invention. The terms used herein in the description of the present invention are for the purpose of describing specific embodiments only, and are not intended to limit the present invention. As used herein, the term "and / or" includes any and all combinations of one or more of t...

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Abstract

The invention relates to a preparation method and a storage method for placenta-derived mesenchymal stem cell exosome, and belongs to the technical field of stem cells and regenerative medicines. Thepreparation method comprises the following steps of: culturing a P3 generation placenta-derived mesenchymal stem cell; after a degrees of fusion reaches 80-90%, sucking the P3 generation placenta-derived mesenchymal stem cell out of a culture medium, and culturing in a DMEM (Dulbecco's modified eagle medium) for 24 h; then, collecting liquid supernatant, and removing cell debris; filtering througha 0.22 [mu] m sterile filtration membrane, taking out a filtrate, carrying out centrifugation at a revolving speed of 13000 r/ min for 60 min, filtering the liquid supernatant through the 0.22 [mu] msterile filtration membrane to obtain high-purity exosome, and finally, carrying out re-suspension by normal saline and centrifugation at a revolving speed of 13000 r/ min for 65 min to obtain the concentrated exosome; and adding the exosome into a cryopreservation liquid containing an SR serum-free medium, glycerol and vitamin C, freezing at a temperature of 80 DEG C below zero for 24h, taking out, and carrying out cryopreservation in liquid nitrogen at a temperature of 196 DEG C below zero. The above preparation method for the placenta-derived mesenchymal stem cell exosome aims at deficiencies in the conventional art, an extraction method is regulated, and the cryopreservation liquid is improved to obtain the high-purity high-density exosome capable of keeping activity.

Description

technical field [0001] The invention relates to the technical field of stem cells and regenerative medicine, in particular to a method for preparing and storing placental mesenchymal stem cell exosomes. Background technique [0002] Mesenchymal stem cells (MSCs) are a type of pluripotent stem cells derived from mesoderm with high renewal and multi-lineage differentiation potential. They can be cultured and expanded in vitro and differentiated into fat cells, bone cells, and liver cells under specific conditions. other tissue cells. Therefore, mesenchymal stem cells are widely used in clinical cell transplantation therapy. However, studies have shown that the proliferation rate of stem cells after transplantation is very low and cannot replace the entire tissue. The main mechanism of cell transplantation therapy is likely to be stem cell exosomes that promote tissue repair rather than simple cell replacement. [0003] Exosomes are bilayer lipid membrane vesicle-like structu...

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Application Information

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IPC IPC(8): C12N5/0775A01N1/02
CPCA01N1/0226C12N5/0665C12N2509/00
Inventor 赵蓝刘小翠李静静江嘉豪孙灿兴何美第
Owner GUANGDONG VITALIFE BIOTECHNOLOGY CO LTD
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