Kit for Preparing a Composition Comprising Fat Cells

• US 20080318317A1
• Filed: 09/20/2006
• Published: 12/25/2008
• Est. Priority Date: 09/20/2005
• Status: Abandoned Application

• Current Assignee: Université de la Réunion
• Original Assignee: Franck Festy, Regis Roche, Université de la Réunion
• Inventors: Roche, Regis, Festy, Franck
• Application Number:
  US12/067,493
• Publication Number:
  US 20080318317A1
• Time in Patent Office: Days
• US Class Current: 435/378
• CPC Class Codes: C12M 45/05 by centrifugation centrifuges in general B04B
• CPC Class Codes: C12M 45/09 by enzymatic treatment
• CPC Class Codes: C12M 47/04 Cell isolation or sorting purging biological preparations of unwanted cells C12N5/0081, determining the presence or kind of microorganism C12Q1/04
• CPC Class Codes: C12N 5/0653 Adipocytes; Adipose tissue

Abstract

A kit for preparing a composition including fat cells includes: a) at least one sterile tube (1) adapted to be centrifuged, including a body (11) and a plug (12) including an element made of elastic and retractable material, the body (11) being hermetically sealed by the plug (12), the plug (12) including an air outlet (13) provided with a filter membrane having a size of pores of less than 0.30 μm; and b) at least one sterile needle including a central channel, one end of which is provided with a device for being fixed to a syringe, and the other end of which is bevelled, the diameter of the central channel being not less than 3 millimeters, preferably not less than 5 millimeters.

Claims

1. A kit for preparing a composition comprising fat cells, said kit comprising:

a) at least one sterile tube (1) adapted to be centrifuged, including a body (11) and a plug (12) comprising an element made of elastic and retractable material, the body (11) being hermetically sealed by the plug (12), said plug (12) including an air outlet (13) provided with a filter membrane having a size of pores of less than 0.30 μ

m;

b) at least one sterile needle, comprising a central channel, one end of which is provided with a device for being fixed to a syringe, and the other end of which is bevelled, the diameter of the central channel being not less than 3 millimeters, preferably not less than 5 millimeters.

2. A kit according to claim 1, characterized in that the plug (12) of the sterile tube (1) comprises at least one sterile puncture area (14) covered with a strippable film (15).

3. A kit according to claim 2, characterized in that the plug (12) of the sterile tube (1) comprises from 2 to 5 puncture areas (14).

4. A kit according to claim 2, characterized in that the plug (12) of the sterile tube (1) comprises more than 5 puncture areas (14).

5. A kit according to claim 1, characterized in that the sterile tube (1) contains an enzyme-based preparation suitable for adipose tissue digestion, said enzyme-based preparation being either in a liquid or in a lyophilized form.

6. A kit according to claim 1, characterized in that the sterile tube (1) does not comprise any enzyme-based preparation.

7. A kit according to claim 6, characterized in that it comprises a container containing an enzyme-based preparation suitable for adipose tissue digestion, said enzyme-based preparation being either in a liquid or in a lyophilized form.

8. A kit according to claim 1, characterized in that it includes 2, 3, 4, 5, 6, 7 or 8 sterile tubes (1).

9. A kit according to claim 1, characterized in that it comprises a hypotonic medium-containing flask suitable for the erythrocyte lysis.

10. A kit according to claim 7, characterized in that said hypotonic medium has a pH value ranging from 7.2 to 7.6.

11. A kit according to claim 1, characterized in that it comprises a flask containing an isotonic medium suitable for cell resuspension.

12. A kit according to claim 1, characterized in that it comprises at least one sterile filtering device provided with a filter membrane having a pore size ranging from 40 μ

m to 200 μ

m.

13. A kit according to claim 12, characterized in that the sterile filtering device further comprises a sterile needle of not less than 16 Gauges.

14. A kit according to claim 1, characterized in that it further comprises at least one needle of not more than 18 Gauges.

15. A kit according to claim 1, characterized in that it further comprises a device (2) for obtaining a composition resulting from the combination of precursor cells with a biological matrix, said device being composed of:

(i) a first syringe (21) intended to be filled with a suitable volume of a biological matrix;

(ii) a second syringe (22) intended to be filled with a precursor cell suspension in an isotonic medium; and

(iii) a pipe (23) connecting the outlet of each syringe (21, 22), said pipe (23) putting the syringes (21) and (22) into fluid communication with each other.

16. A method for preparing a composition comprising fat cells, said method comprising following steps of:

a) introducing an adipose tissue suspension resulting from a liposuction or a lipectomy into a tube (1) such as defined in claim 1;

b) allowing an enzymatic digestion of the adipose tissue suspension to proceed in the tube (1) using an enzyme-based preparation containing at least one protease;

c) centrifuging the suspension after the enzymatic digestion;

d) collecting the centrifuged suspension fractions containing the interesting cells;

d1) collecting the adipocyte precursor cell (AP) fraction in the pellet located in the bottom portion, of the tube (1);

d2) if necessary, collecting the mature adipocyte (MA) fraction recovered in the upper part of the centrifuged adipose tissue suspension;

e) incubating the (AP) fraction in a hypotonic solution so as to lyse erythrocytes; and

f) preparing a fat cell composition from one of the following cell fractions;

f1) the (AP) fraction of adipocyte precursor cells such as obtained at the end of step e);

f2) the (MA) fraction of mature adipocytes such as obtained in step d1); and

f3) a combination of cells of the (AP) fraction and cells of the (MA) fraction.

17. A method according to claim 16, characterized in that in step f), when the combination is made with both (AP) fraction cells and (MA) fraction cells, the cell ratio (AP):

(MA) does vary from 1;

99 to 99;

1.

18. A sterile tube (1) adapted to be centrifuged, including a body (11) and a plug (12) comprising an element made of elastic and retractable material, the body (11) being hermetically sealed by the plug (12), said plug (12) including an air outlet (13) provided with a filter membrane having a size of pores of less than 0.30 μ

m.

19. A sterile tube (1) according to claim 18, characterized in that the plug (12) of the sterile tube (1) comprises at least one sterile puncture area (14) covered with a strippable film (15).

20. A sterile tube (1) according to claim 18, characterized in that the plug (12) of the sterile tube (1) comprises at least two puncture areas (14).

21. A sterile tube (1) according to claim 18, characterized in that it contains an enzyme-based preparation suitable for adipose tissue digestion, said enzyme-based preparation being either in a liquid or in a lyophilized form.