Selective lysing of cells using ultrasound
View Patent ↗Described are embodiments that employ ultrasonic energy to selectively lyse larger adipose cells in a suspension containing adipose cells of different sizes resulting in a suspension in which the only viable cells are the small adipose cells and stem cells. Embodiments provide for generating an acoustic standing wave field of sufficient intensity and proper geometry, that high shear stress is induced on the cell membranes of cells larger than a predetermined size. The remaining small adipose cells can be physically separated from the suspension after the suspension is subjected to the acoustic standing wave field.
1. A method for lysing adipose cells of diameters equal to or greater than a predetermined diameter, the method comprising:
generating an acoustic standing wave field, wherein the geometry of the acoustic standing wave field is such that one half of the wavelength of the acoustic standing wave field is less than or equal to the predetermined diameter, and wherein the intensity of the acoustic standing wave field is sufficient to cause lysing of adipose cells of diameters equal to or greater than the predetermined diameter; and
subjecting a mixture of adipose cells and stem cells to the acoustic standing wave field to generate a lysed mixture, wherein the mixture of adipose cells and stem cells includes at least some adipose cells having a diameter equal to or greater than the predetermined diameter.
2. The method of claim 1 , wherein the subjecting results in selective lysing of adipose cells equal to or greater than the predetermined diameter compared to cells of a smaller diameter.
3. The method of claim 1 , wherein the acoustic standing wave field is an ultrasonic standing wave field.
4. The method of claim 1 , further comprising:
processing the lysed mixture to prepare at least a portion of the lysed mixture for injection into a patient.
5. The method of claim 4 , wherein the processing comprises:
a separation process that separates intact cells from other components of the lysed mixture, the separation process comprising at least one of: filtration, centrifugation, and settling.
6. The method of claim 1 , wherein the subjecting comprises holding a batch of the mixture in a chamber that includes the acoustic standing wave field for a preset dwell time.
7. The method of claim 1 , wherein the subjecting comprises:
flowing the mixture through a chamber that includes the acoustic standing wave field.
8. The method of claim 1 , wherein the method further comprises lysing adipose cells of diameters equal to or greater than a second predetermined diameter that is smaller than the first predetermined diameter by:
generating a second acoustic standing wave field, wherein the geometry of the second acoustic standing wave field is such that one half of the wavelength of the second acoustic standing wave field is less than or equal to the second predetermined diameter, and wherein the intensity of the second acoustic standing wave field is sufficient to cause lysing of adipose cells of diameters equal to or greater than the second predetermined diameter; and
subjecting the lysed mixture to the second acoustic standing wave field, wherein the lysed mixture includes at least some adipose cells having a diameter equal to or greater than the second predetermined diameter.
9. A method for lysing adipose cells of diameters equal to or greater than a predetermined diameter, the method comprising:
generating an acoustic standing wave field, wherein the geometry of the acoustic standing wave field is such that one half of the wavelength of the acoustic standing wave field is less than or equal to the predetermined diameter, the wavelength of the acoustic standing wave field falling within a range of 0.5 mm to 2.5 mm, and wherein the intensity of the acoustic standing wave field is sufficient to cause lysing of adipose cells of diameters equal to or greater than the predetermined diameter; and
subjecting a mixture of adipose cells and stem cells to the acoustic standing wave field to generate a lysed mixture, wherein the mixture of adipose cells and stem cells includes at least some adipose cells having a diameter equal to or greater than the predetermined diameter.
10. The method of claim 9 , wherein the subjecting results in selective lysing of adipose cells equal to or greater than the predetermined diameter compared to cells of a smaller diameter.
11. The method of claim 9 , wherein the acoustic standing wave field is an ultrasonic standing wave field.
12. The method of claim 9 , further comprising:
processing the lysed mixture to prepare at least a portion of the lysed mixture for injection into a patient.
13. The method of claim 12 , wherein the processing comprises:
a separation process that separates intact cells from other components of the lysed mixture, the separation process comprising at least one of: filtration, centrifugation, and settling.
14. The method of claim 9 , wherein the subjecting comprises holding a batch of the mixture in a chamber that includes the acoustic standing wave field for a preset dwell time.
15. The method of claim 9 , wherein the subjecting comprises:
flowing the mixture through a chamber that includes the acoustic standing wave field.
16. The method of claim 9 , wherein the method further comprises lysing adipose cells of diameters equal to or greater than a second predetermined diameter that is smaller than the first predetermined diameter by:
generating a second acoustic standing wave field, wherein the geometry of the second acoustic standing wave field is such that one half of the wavelength of the second acoustic standing wave field is less than or equal to the second predetermined diameter, and wherein the intensity of the second acoustic standing wave field is sufficient to cause lysing of adipose cells of diameters equal to or greater than the second predetermined diameter; and
subjecting the lysed mixture to the second acoustic standing wave field, wherein the lysed mixture includes at least some adipose cells having a diameter equal to or greater than the second predetermined diameter.