Brown-like fat exosome and engineering production method and application thereof

A brown fat, production method technology, applied in the field of cell engineering and gene recombination, can solve the problem of limited source of brown fat exosomes

Active Publication Date: 2019-11-22
FOURTH MILITARY MEDICAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] In view of this, the purpose of the present invention is to provide an engineering production method of brown fat exosomes, which solves the problem of limited source of brown fat exosomes, and transforms unnecessary white fat stem cells such as obesity into brown fat stem cells , realize engineering production on the basis of individual specificity

Method used

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  • Brown-like fat exosome and engineering production method and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0055] 1) Obtain subcutaneous fat stem cells (white fat stem cells)

[0056] Under sterile conditions, remove the fascia from the fat tissue. Transfer the removed adipose tissue to a petri dish containing sterile PBS, wash it 2 to 3 times, take it out and put it in a petri dish and cut it into pieces. After the tissue is minced, transfer the tissue to a centrifuge tube containing 0.1% collagenase I, under the condition of 5% CO 2 , Digest in an incubator at 37°C, and shake once every 10 minutes until the fat tissue is completely digested into a suspension. Add an equal amount of complete medium DMEM (containing 1% penicillin-streptomycin, 10% fetal bovine serum) to further dilute the suspension so that the filtrate can be better filtered out by a 200-mesh steel sieve. The filtrate was transferred to a centrifuge tube and centrifuged at 1000r / min for 5min at room temperature. Take the supernatant and the upper incompletely digested adipose tissue, add the complete medium DME...

Embodiment 2

[0069] The brown fat-like exosomes prepared in Example 1 were injected into obese mice (100 μg / mouse, once a week) through the tail vein, and the obese mice group without exosome injection was used as a control, and the same method was used to raise 3 After one week, the body weight of mice injected with brown fat-like exosomes dropped significantly from an average of 35.5 g to 31.2 g. There was no significant change in body weight of mice in the control group.

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Abstract

The invention provides a brown-like fat exosome and an engineering production method and application thereof, and belongs to the technical field of cell engineering and gene recombination. The engineering production method of the brown-like adipose exosome comprises the following steps: separating adipose-derived stem cells from adipose tissues; constructing a virus expression vector for simultaneously expressing PGC1a and the PRDM16; packaging the virus expression vector into a virus; infecting the adipose-derived stem cells with the virus; culturing the adipose-derived stem cells infected with the virus, and collecting the brown-like fat exosome. Through the genetic modification method, pGC1a and PRDM16 genes are introduced into adipose-derived stem cells to activate the expression of aplurality of brown fat-related genes in the adipose-derived stem cells, the adipose-derived stem cells are modified into brown adipose-derived stem cells, and then the brown-like fat exosome is generated. The brown-like fat exosome has characteristics of brown fat exosomes, and a potential treatment means is provided for metabolic diseases such as obesity.

Description

technical field [0001] The invention belongs to the technical field of cell engineering and gene recombination, and in particular relates to a brown fat-like exosome and its engineering production method and application. Background technique [0002] Obesity is the most important cause of metabolic diseases. With the change of human life style, the incidence of obesity increases year by year, increasing the risk of various diseases including cardiovascular disease, tumor, diabetes and so on. Up to now, there is still a lack of ideal drug treatment. [0003] The main function of brown fat is to consume energy and produce heat, and it is an important tissue for the body to fight obesity. But as obesity increases and age increases, brown adipose tissue gradually decreases. Brown fat exosomes, as vesicles secreted by brown fat, have functions similar to brown fat and are potential therapeutic drugs for metabolic diseases. However, as a kind of non-renewable resource, brown f...

Claims

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Application Information

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IPC IPC(8): C12N15/867C12N5/10A61K35/35A61P3/04A61P1/16A61P17/02
CPCA61K35/35A61P1/16A61P3/04A61P17/02C12N5/0667C12N15/86C12N2740/15043
Inventor 杨国栋周雪莹袁丽君孙汶齐李者龙邢长洋赵联璧刘云楠王辰
Owner FOURTH MILITARY MEDICAL UNIVERSITY
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