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
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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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