Fat extract without additive components as well as preparation method and application of fat extract
A fat extraction, additive-free technology, applied in the biological field, can solve the problems of unsatisfactory anti-aging effect
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Embodiment 1
[0109] Embodiment 1 fat extract preparation
[0110] Fat was obtained from volunteers with informed consent. The fat extraction method is as follows:
[0111] (1) Take the fat obtained by suction or surgical resection, shred it and rinse it with normal saline for 3 times.
[0112] (2) Take the rinsed adipose tissue, put it in a 50ml centrifuge tube (about 30-50ml per tube), put it into a centrifuge and centrifuge at 1200rpm for 3 minutes to obtain a layered mixture.
[0113] (3) For the layered mixture, drain the excess liquid at the bottom and the uppermost layer of fat, and collect the middle layer (ie, the fat layer containing fat cells).
[0114] (4) For the middle layer, use two 10ml injection syringes to connect the three-way pipe to beat about 60-120 times, thereby performing mechanical emulsification, and obtaining a mechanically emulsified fat mixture (also called nanofat).
[0115] (5) Put the mechanically emulsified fat mixture (which can be combined or not) into...
Embodiment 2
[0117] The influence of embodiment 2 fat extracts on human skin fibroblast proliferation
[0118] Human skin fibroblasts were isolated from neonatal foreskin tissue, then inoculated in high-glucose DMEM medium containing 10% fetal bovine serum, and passaged once every 3 days. The 3rd and 4th passage cells were used in the experiment.
[0119] Human skin fibroblasts were inoculated in a 96-well plate at a density of 1,000 cells per well, and fat extracts of different concentrations (1%-5%) were added, and 6 secondary wells were set for each concentration, and used after culturing for 72 hours. The CCK8 kit was used to detect cell proliferation and determine the OD value. The experiment was repeated three times to obtain the average value and standard deviation.
[0120] result:
[0121] Human skin fibroblasts were cultured with different concentrations of fat extracts, CCK-8 detection showed that the number of cells added in the extract group was significantly higher than that...
Embodiment 3
[0123] Example 3 Effect of Fat Extraction on Anti-aging and Oxidative Stress of Skin Tissue
[0124] Treat skin fibroblasts with fat extracts of different concentrations (1%-5%) for 24 hours, ultraviolet rays (100mJ / cm 2 ) after irradiation, the content of reactive oxygen species (ROS) in the cells was detected by flow cytometry.
[0125] After 48 hours after irradiation, the cell cycle was detected by flow cytometry.
[0126] After 72 hours of irradiation, cell proliferation was detected with a CCK8 kit, and senescent cells were stained with beta-gal and phalloidin to detect the degree of aging. In addition, the RNA of the cells was extracted, and the expression of type I collagen was detected by RT-PCR technique.
[0127] result:
[0128] 3.1 ROS content
[0129] Human skin fibroblasts were cultured with fat extracts at different concentrations, intracellular ROS staining was performed immediately after ultraviolet light irradiation, and the accumulation of intracellular...
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