ISOLATION OF STEM CELLS FROM ADIPOSE TISSUE BY ULTRASONIC CAVITATION, AND METHODS OF USE

• US 20150159151A1
• Filed: 06/26/2013
• Published: 06/11/2015
• Est. Priority Date: 06/26/2012
• Status: Abandoned Application

• Current Assignee: Amberdale Enterprises Pty. Ltd., Tavid Pty Ltd.
• Original Assignee: Amberdale Enterprises Pty. Ltd., Rusty Property Holdings Pty Ltd., Tavid Pty Ltd.
• Inventors: Bright, Ralph, Bright, Pelin, Hansen, Bruce, Thomas, Wayne
• Application Number:
  US14/409,229
• Publication Number:
  US 20150159151A1
• Time in Patent Office: Days
• US Class Current: 1/1
• CPC Class Codes: A61K 35/35 Fat tissue; Adipocytes; Stromal cells; Connective tissues adipose-derived stem cells A61K35/28; collagen A61K38/39
• CPC Class Codes: A61P 11/00 Drugs for disorders of the respiratory system
• CPC Class Codes: A61P 19/00 Drugs for skeletal disorders
• CPC Class Codes: A61P 19/02 for joint disorders, e.g. arthritis, arthrosis
• CPC Class Codes: A61P 21/00 Drugs for disorders of the muscular or neuromuscular system
• CPC Class Codes: A61P 21/04 for myasthenia gravis
• CPC Class Codes: A61P 25/00 Drugs for disorders of the nervous system
• CPC Class Codes: A61P 27/00 Drugs for disorders of the senses
• CPC Class Codes: A61P 29/00 Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
• CPC Class Codes: A61P 37/02 Immunomodulators
• CPC Class Codes: C12N 1/066 by physical methods
• CPC Class Codes: C12N 13/00 Treatment of microorganisms or enzymes with electrical or wave energy, e.g. magnetism, sonic waves
• CPC Class Codes: C12N 2521/10 Sound, e.g. ultrasounds
• CPC Class Codes: C12N 5/0667 Adipose-derived stem cells [ADSC]; Adipose stromal stem cells

Abstract

In one embodiment, the present invention relates to a non-enzymatic method for isolating stem cells from adipose tissue, wherein the method comprises treating adipose tissue with ultrasonic cavitation to break up the adipose tissue and lyses mature adipocytes, resulting in a stromal vascular fraction containing viable stromal/stem cells.

Claims

1. A method of isolating a stromal vascular fraction comprising viable stromal/stem cells from adipose tissue, the method comprising treating the adipose tissue with ultrasonic cavitation for a time, amplitude and cycle sufficient to break up the tissue and lyse mature adipocytes while maintaining the viability of the stromal/stem cells.

2. The method according to claim 1 wherein the temperature of the adipose tissue during ultrasonic cavitation is maintained at a temperature that ensures the viability of the stromal/stem cells.

3. The method according to claim 1 wherein the temperature of the adipose tissue does not exceed about 43°

C. to about 45°

C.

4. The method according to claim 1 wherein the adipose tissue is treated with ultrasonic cavitation for about 10 seconds to about 10 min with an ultrasonic device set at amplitude about 20 to about 90% and cycle about 0.2 to about 0.9.

5. The method according to claim 1 wherein the time, amplitude and cycle are selected such that the temperature of the adipose tissue does not exceed about 43°

C. to about 45°

C.

6. The method according to claim 1 wherein the adipose tissue is treated with ultrasonic cavitation for about 1 minute and 30 seconds to about 1 minute and 40 seconds with an ultrasonic probe set at amplitude about 50% and cycle about 0.4 to about 0.5.

7. The method according to claim 1 wherein the lipid layer resulting from the ultrasonic cavitation is removed before addition of an isotonic solution.

8. A stromal vascular fraction comprising viable stromal/stem cells when isolated by the method according to claim 1.

9. A method of treating osteoarthritis, a joint-related inflammatory disorder, an inflammatory arthritis disorder, soft-tissue damage or tears, a cartilage disorder, a bone disorder, an auto-immune disorder, muscle dystrophy, chronic fatigue, a lung disorder, a lung inflammatory disorder, a nervous system disorder, a spinal disorder or a neurological disorder in a subject comprising administering to the subject a stromal vascular fraction comprising viable stromal/stem cells according to claim 8.

10. A method of treating osteoarthritis, a joint-related inflammatory disorder, an inflammatory arthritis disorder, soft-tissue damage or tears, a cartilage disorder, a bone disorder, an auto-immune disorder, muscle dystrophy, chronic fatigue, a lung disorder, a lung inflammatory disorder, a nervous system disorder, a spinal disorder or a neurological disorder in a subject comprising administering to the subject stromal vascular fraction comprising viable stromal/stem cells, wherein the stromal vascular fraction comprising viable stromal/stem cells has been isolated by the method according to claim 1.

11-14. -14. (canceled)

15. A method of isolating a stromal vascular fraction comprising viable stromal/stem cells from adipose tissue comprising treating the adipose tissue with ultrasonic cavitation for about 1 minute and 30 seconds to about 1 minute and 40 seconds with an ultrasonic device set at an amplitude and cycle that ensures that the temperature of the adipose tissue does not exceed about 43°

C. to about 45°

C.

16. A method of isolating a stromal vascular fraction comprising viable stromal/stem cells from adipose tissue comprising treating the adipose tissue with an ultrasonic device set at an amplitude of about 50%, wherein the device is set at a cycle and applied for a time that ensures that the temperature of the adipose tissue does not exceed about 43°

C. to about 45°

C.

17. A method of isolating a stromal vascular fraction comprising viable stromal/stem cells from adipose tissue comprising treating the adipose tissue with an ultrasonic device set at a cycle about 0.4 to about 0.5, wherein the device is set at an amplitude and applied for a time that ensures that the temperature of the adipose tissue does not exceed about 43°

C. to about 45°

C.

18. A method of isolating a stromal vascular fraction comprising viable stromal/stem cells from adipose tissue comprising treating the adipose tissue with ultrasonic cavitation for about 1 minute and 30 seconds to about 1 minute and 40 seconds with an ultrasonic probe set at amplitude about 50% and cycle about 0.4 to about 0.5.

19. A method for maintaining viability of stromal/stem cells in the stromal vascular fraction isolated from adipose tissue by ultrasonic cavitation, wherein time, amplitude and cycle of ultrasonic cavitation are selected such that the stromal/stem cells are viable after 48 hours cell culture.