A fat graft enriched with highly active fat granule cells and adipose stem cells, its preparation method and application
A technology of fat stem cells and fat granules, applied in the field of fat transplantation, can solve the problems of only 10% gel yield, insufficient fat storage, and low absorption rate of SVF-gel
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Embodiment 1
[0041] The purpose of this example is to select the most suitable liposuction method.
[0042] The fat extraction method of the present embodiment is as follows:
[0043] Negative pressure liposuction machine: use the currently commonly used machine to assist the negative pressure liposuction machine to extract fat;
[0044] Manual extraction: Use a 10mL or 20mL syringe for manual negative pressure liposuction. The extracted fat is left standing at 4°C under sterile conditions to remove water to obtain fat (granular fat). Use 3-5 holes for liposuction with a diameter of 1.2-2mm Needle, liposuction syringe is best to use 10mL or 20mL syringe.
[0045] The fat treatment process and result records of the two groups are shown in Table 1, as follows:
[0046] Table 1
[0047]
[0048]
[0049] The results of the two groups showed that compared with manual extraction, the fat from machine-assisted negative pressure liposuction needs to increase the number of centrifugation ...
Embodiment 2
[0051] The purpose of this example is to examine the effect of mechanical treatment on the viability of cells in fat.
[0052] Each group of fat preparation process of the present embodiment is as follows:
[0053] Control group: common fat treated by centrifugation to remove water;
[0054] Group A: (1) manual negative pressure liposuction with 10mL or 20mL syringe, the extracted fat was left to stand under sterile conditions at 4°C to remove water to obtain fat particles; the aperture of the liposuction needle used was 1.2-2mm, The number of holes in the needle is 3 to 5;
[0055] (2) Add physiological saline twice the volume of the fat particles in step (1), shake and mix well, and centrifuge at 300-500g for 3 minutes. After centrifugation, the fat is divided into the upper fat layer and the lower water layer, discard the lower water layer and tube The basal erythrocytes are precipitated to obtain the fat layer;
[0056] (3) The fat layer obtained in step (2) is divided ...
Embodiment 3
[0078] The purpose of this example is to detect the effect of mechanical treatment on the activity of adipose stem cells in fat grafts.
[0079] The specific operation is as follows:
[0080] (1) Digest and collect the SVF-cells obtained from the four groups of fats in Example 2, remove red blood cells, and press 2×10 6 Cells / mL were resuspended and cultured in 6-well plate culture flasks, 1 mL / well, and about 2×10 cells were inoculated in each well. 6 cells. After 24 hours of attachment, observe the general adhesion of the cells in the four groups of flasks.
[0081] (2) Suck off the supernatant in the culture flask, add normal saline to wash twice, add 0.25% trypsin to digest in each bottle, neutralize and digest, centrifuge at 400g for 3min, discard the supernatant to obtain cell pellet;
[0082] (3) Prepare the cell pellet as a cell suspension, add 0.4% trypan blue to the cell suspension at a volume of 9:1 for staining (the volume ratio of cell suspension: 0.4% trypan b...
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