Isolation of stem cells from adipose tissue by ultrasonic cavitation, and methods of use

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.

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 . (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.