Rapid mincing of adipose tissues to isolate live cells in vitro

1. A method for isolating live, stromal vascular fraction (SVF) cells in vitro from adipose tissues, comprising the steps as follows:

step (a) providing an adipose tissue;

step (b) arranging the adipose tissue into an enclosed vessel of a cutting device, wherein the cutting device is a disposable homogenizer comprising a power unit;

step (c) applying an acting force on the enclosed vessel by the cutting device to enable a preset cutting tool in the enclosed vessel to implement rapid and homogeneous mincing on the adipose tissue in a closed environment, thereby obtaining a homogenized adipose tissue, wherein a relative centrifugal force of the cutting tool is 16-25×g;

step (d) adding a reagent into the homogenized adipose tissue to implement a hydrolysis reaction;

step (e) filtering and centrifugally separating the homogenized adipose tissue treated via the hydrolysis reaction; and

step (f) removing a supernate after the centrifugal separation so as to obtain a cell pellet, which contains isolated stromal vascular fraction (SVF) cells.

2. The method for isolating live, stromal vascular fraction (SVF) cells in vitro from adipose tissues according to claim 1 , wherein a step of cleaning the adipose tissue via a phosphate buffer solution is included before step (a).

3. The method for isolating live, stromal vascular fraction (SVF) cells in vitro from adipose tissues according to claim 1 , wherein the adipose tissue in step (a) is a lump adipose tissue obtained from surgery excision of a mammal.

4. The method for isolating live, stromal vascular fraction (SVF) cells in vitro from adipose tissues according to claim 3 , wherein the mammal is a human being.

5. The method for isolating live, stromal vascular fraction (SVF) cells in vitro from adipose tissues according to claim 1 , wherein the disposable homogenizer comprises a disposable enclosed vessel, and the inside of the disposable enclosed vessel is provided with a cutting tool.

6. The method for isolating live, stromal vascular fraction (SVF) adipose tissues according to claim 5 , wherein the homogeneous mincing in step (c) is performed by arranging 1 g to 6 g of adipose tissues into the disposable enclosed vessel with a volume of 15 ml to implement rapid and homogeneous mincing.

7. The method for isolating live, stromal vascular fraction (SVF) cells in vitro from adipose tissues according to claim 6 , wherein the homogeneous mincing in step (c) is performed by arranging 3 g of adipose tissues into the disposable enclosed vessel with a volume of 15 ml to implement rapid and homogeneous mincing.

8. The method for isolating live, stromal vascular fraction (SVF) cells in vitro from adipose tissues according to claim 7 , wherein an acting time of the power unit is 3-10 minutes.

9. The method for isolating live, stromal vascular fraction (SVF) cells in vitro from adipose tissues according to claim 1 , wherein the reagent in step (d) is selected from the group consisting of a trypsin, a dispase, a gelatase, a hyaluronidase, a collagenase type I, a collagenase type IV and combinations thereof.

10. The method for isolating live, stromal vascular fraction (SVF) cells in vitro from adipose tissues according to claim 1 , wherein the hydrolysis reaction in step (d) is implemented in a constant temperature hybridization reaction oven for the whole process under conditions of 4-45 DEG C., 5 rpm-50 rpm and 0.5 h-24 h.

11. The method for isolating live, stromal vascular fraction (SVF) cells in vitro from adipose tissues according to claim 10 , wherein the preferred conditions are 37 DEG C., 15 rpm and 8 h.

12. The method for isolating live, stromal vascular fraction (SVF) cells in vitro from adipose tissues according to claim 1 , wherein the conditions for centrifugation in step (e) are 400×g and 10 minutes.

13. The method for isolating live, stromal vascular fraction (SVF) cells in vitro from adipose tissues according to claim 1 , wherein step (e) comprises a step of moving a filtered fluid obtained after filtering the homogenized adipose tissue treated via the hydrolysis reaction into a centrifugal tube, so as to implement centrifugal separation.

14. The method for isolating live, stromal vascular fraction (SVF) cells in vitro from adipose tissues according to claim 1 , wherein after step (f), performing a step (g) of adding the phosphate buffer solution into the cell pellet, cleaning the cell pellet, and then centrifuging again so as to remove a supernate comprising blood and the reagent to obtain a cleaned cell pellet.

15. The method for isolating live, stromal vascular fraction (SVF) cells in vitro from adipose tissues according to claim 14 , wherein after being cleaned in step (g), a step (h) of arranging the cell pellet into a culture medium for culturing so as to obtain amplified cells.

16. The method for isolating live, stromal vascular fraction (SVF) cells in vitro from adipose tissues according to claim 15 , wherein the culture medium comprises a basal culture medium (IMDM), a serum additive and a fibroblast growth factor-2 (FGF-2).

17. The method for isolating live, stromal vascular fraction (SVF) cells in vitro from adipose tissues according to claim 16 , wherein the serum additive is a fetal bovine serum with a volume percentage concentration of 2% to 10%, and a concentration of the FGF-2 is 1 ng/ml to 20 ng/ml.

18. The method for isolating live, stromal vascular fraction (SVF) cells in vitro from adipose tissues according to claim 15 , wherein the amplified cell is an adipose tissue-derived stem cell which is substantially undifferentiated.

19. The method for isolating live, stromal vascular fraction (SVF) cells in vitro from adipose tissues according to claim 18 , wherein the adipose tissue-derived stem cell at least shows a cell surface antigen CD90, or CD 105 or a combination thereof, and does not show CD45.

20. A method for cell therapy, regenerative medicine or cell and tissue engineering, comprising;

making an adipose tissue-derived stem cell according to the method of claim 19 ; and

applying the adipose tissue-derived stem cells singly or in combination as a material for cell therapy, regenerative medicine or cell and tissue engineering.

21. A method for treating one or a combination of degenerated diseases, tissue damage repair, organ regeneration or tissue regeneration, comprising;

making an adipose tissue-derived stem cell according to the method of claim 18 ;

and

preparing medicinal compositions for cell therapy from the adipose tissue-derived stem cell.

22. A method for establishing a cell bank, comprising;

making an amplified adipose tissue-derived stem cells by the method according to claim 18 ; and

implementing cryopreservation on the amplified adipose tissue-derived stem cell.

23. The method for isolating live, stromal vascular fraction (SVF) cells in vitro from adipose tissues according to claim 18 , wherein a step (i′) of implementing induced differentiation on the amplified adipose tissue-derived stem cell so as to obtain a cell differentiated from the amplified adipose tissue-derived stem cell is included after step (h).

24. The method for isolating live, stromal vascular fraction (SVF) cells in vitro from adipose tissues according to claim 23 , wherein the cells differentiated from the amplified adipose tissue-derived stem cells include one of an osteogenic cell, an adipose cell or a cartilage cell.