Stem cell composition for chest fat filling and application thereof
A technology of fat stem cells and stem cells, applied in drug delivery, tissue regeneration, prostheses, etc., can solve problems such as lifespan and many side effects, breast hardening, easy absorption, etc., to promote the survival and growth of stem cells in the body, reduce body fat Tissue reduction, breasts feel natural effect
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Embodiment 1
[0035]Example 1 Adipose stem cell preparation
[0036] 1. Primary culture of adipose-derived stem cells
[0037] Cut the washed adipose tissue into about 1mm 3 Add the small pieces of type I collagenase (concentration: 1mg / ml) equal to the volume of adipose tissue and mix well, put them in a constant temperature shaker at 37°C for digestion for 1h; filter to remove larger tissue pieces after digestion, and transfer the filtered liquid Transfer to a centrifuge tube, centrifuge at 2000rpm for 10min, wash the cells and remove the cleaning solution, resuspend and mix the cells with 10ml adipose-derived stem cell medium (low-sugar DMEM), inoculate in a T75 culture flask, and store at 37°C, CO 2 The culture was started at a concentration of 5%.
[0038] The primary cells are in the culture bottle for 2 to 3 days, the cells begin to precipitate and adhere to the wall, the color of the culture solution changes from pink to light yellow, then the medium can be changed, the culture me...
Embodiment 2
[0041] Embodiment 2 fibroblast preparation
[0042] Primary cultured skin tissue: Rinse the skin 3 times with PBS containing penicillin and streptomycin to remove the subcutaneous connective tissue as much as possible; cut the skin into small pieces, spread them on a plate with the dermis facing down, add Dispase II and leave overnight at 4°C; Separate the epidermis and dermis the next day, cut the dermis into small pieces, inoculate them in 35mm culture dishes, and place them at 37°C and 5% CO 2 Cultivate in a constant temperature incubator for 1 h, add a small amount of DMEM / F12 medium containing 10% fetal bovine serum (the ratio of DMEM medium to F12 medium is 3:1), continue culturing the next day, and change the medium every 3 days thereafter.
[0043] Subculture: Digest with 0.25% trypsin-0.02% EDTA for about 1 min, observe under the microscope that the cells shrink and become round, and the intercellular space increases, then add DMEM / F12 medium (3:1) containing 10% feta...
Embodiment 3
[0048] Example 3 Preparation of CGF
[0049] Place two blood collection tubes containing 5ml of blood (without adding anticoagulant) symmetrically into the MEDIFUGE centrifuge, and perform differential centrifugation. The program of the differential centrifugation is: acceleration for 30 seconds; centrifugation at 2700 rpm for 2 minutes Centrifuge at 2400 rpm for 4 minutes; centrifuge at 2700 rpm for 4 minutes; centrifuge at 3000 rpm for 3 minutes; decelerate for 33 seconds. After centrifugation, the blood sample is divided into three layers: the upper layer is serum, the middle layer is CGF, and the lower layer is red blood cells. CGF, shredded fibrin clot, collected for later use.
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