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Method for preparing adipose-derived stem cell exosomes, adipose-derived stem cell exosomes and application thereof

A technology of fat stem cells and exosomes, applied in the field of bioengineering, can solve the problem of weak proliferation of endometrial microvascular endothelial cells

Active Publication Date: 2019-06-28
SHANGHAI NINTH PEOPLES HOSPITAL AFFILIATED TO SHANGHAI JIAO TONG UNIV SCHOOL OF MEDICINE
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Problems solved by technology

In the research work, it was found that exosomes from adipose-derived stem cells had a weak effect on promoting the proliferation of endometrial microvascular endothelial cells. Therefore, how to enhance the angiogenesis effect of adipose-derived stem cell exosomes obtained in vitro is an important problem that needs to be solved.

Method used

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  • Method for preparing adipose-derived stem cell exosomes, adipose-derived stem cell exosomes and application thereof
  • Method for preparing adipose-derived stem cell exosomes, adipose-derived stem cell exosomes and application thereof
  • Method for preparing adipose-derived stem cell exosomes, adipose-derived stem cell exosomes and application thereof

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Experimental program
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Embodiment 1

[0054] 1. Isolation and Identification of Adipose Stem Cells: Adipose tissue is the abdominal or thigh fat waste obtained by liposuction under sterile conditions from the Plastic Surgery Department of the Ninth People's Hospital Affiliated to Shanghai Jiaotong University School of Medicine. The adipose tissue was centrifuged at 1500rpm for 5 minutes , discard the upper layer of fat and the lower blood cell layer, take the middle fat layer, wash the fat with PBS for 3 to 5 times, until the cleaning solution is clear; the mixture obtained from the last washing is centrifuged at 1500rpm for 5 minutes, absorb the fat, and record the fat volume; Use digestive enzymes at a ratio of 1:1 to digest, digest at 37°C constant temperature shaker at 150-280rpm for 20-60 minutes; after digestion, centrifuge at 2000rpm for 5 minutes, discard the upper fat layer and the digested fat layer, leaving a layer of dry cell sediment and A small amount of digestion solution was added to terminate with ...

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Abstract

The invention provides a method for preparing adipose-derived stem cell exosomes, the adipose-derived stem cell exosomes and application thereof and belongs to the technical field of bioengineering. The preparing method comprises the steps: 1, mixing adipose-derived stem cells with an electrotransfection working solution to obtain a mixed solution, and mixing the mixed solution with a CTF1 plasmidto obtain a pre-transfection object; 2, conductng electric shock on a pre-transfection material obtained in the step 1 to obtain adipose stem cells transfected with a CTF1 gene; 3, carrying out conventional culture on the adipose stem cells transfected with the CTF1 gene obtained in the step 2, and collecting the adipose-derived stem cell exosomes. The adipose-derived stem cell exosomes preparedby the method can remarkably promote proliferation of endometrial microvascular endothelial cells.

Description

technical field [0001] The invention relates to the technical field of bioengineering, in particular to a method for preparing exosomes from adipose stem cells, adipose stem cells and applications thereof. Background technique [0002] Angiogenesis establishes a rich blood circulation for the endometrium at the embryo attachment site, providing a material basis for embryo implantation. The richer the subendometrial blood flow, the better the endometrial receptivity, and the more conducive to embryo implantation. Increasing endometrial blood supply and improving endometrial receptivity by intervening in the process of endometrial angiogenesis is an important means to improve the embryo implantation rate. Aspirin is considered to increase the blood perfusion of uterus and ovary, thereby increasing the embryo implantation rate and clinical pregnancy rate, so it is widely used clinically, but its efficacy is still controversial. Production rate, and reduce the miscarriage rate...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/10C12N15/85A61K35/28A61P15/00
Inventor 林嘉盈黄嘉膂蔡仁飞匡延平
Owner SHANGHAI NINTH PEOPLES HOSPITAL AFFILIATED TO SHANGHAI JIAO TONG UNIV SCHOOL OF MEDICINE
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