Separation, dissociation and/or disaggregation of cells using shockwaves or mechanical impacts
Methods provided by the present disclosure utilize extracorporeal shockwaves, mechanical impacts and/or principles of lithotripsy to break up a tissue sample into smaller fragments—clusters of cells and/or single cells—after which a desired cellular fraction can be isolated from the sample. Devices provided by the present disclosure deploy focused and/or directed shockwaves, and/or focused and directed mechanical impacts, to break apart a tissue sample. The devices maintain the sample in a sterile, closed environment during exposure to the shockwaves or mechanical impacts. Therefore, the shockwaves and/or mechanical impacts are generated outside of a closed device and are transmitted through one or more walls of the device into its interior, where the sample is located.
1. A method of isolating a stromal vascular fraction from tissue, comprising:
placing a tissue sample into a processing container of a tissue processing device;
subjecting the tissue sample to force from mechanical impacts to break down the tissue; and
separating the stromal vascular fraction from the tissue;
wherein:
the mechanical impacts are generated by an impact arm that makes physical contact with the processing container; and
the impact arm does not physically contact the tissue.
2. The method of claim 1 , wherein the processing device comprises:
a platform to which the processing container is secured; and
a first motor that drives up and down articulation of the impact arm.
3. The method of claim 1 , wherein the processing container is a cartridge.
4. The method of claim 1 , wherein the force from the mechanical impacts is delivered to the tissue through a wall of the processing container.
5. The method of claim 1 , wherein the separating comprises allowing the tissue to separate from an aqueous layer, the aqueous layer comprising the stromal vascular fraction.
6. The method of claim 1 , wherein the stromal vascular fraction is isolated from the tissue in 30 minutes or less.
7. The method of claim 1 , wherein the stromal vascular fraction comprises stem cells, growth factors and progenitor cells.
8. The method of claim 1 , wherein the tissue is selected from adipose tissue, brain tissue, pharyngeal tissue, laryngeal tissue, heart tissue, arterial tissue, muscle tissue, liver tissue, gall bladder tissue, kidney tissue, small intestinal tissue, large intestinal tissue, lymph node tissue, lung tissue, spleen tissue, bone marrow tissue, stomach tissue, venous tissue, pancreatic tissue, urinary bladder tissue, bone, teeth, dentin tissue, gum tissue, skin tissue, pineal gland tissue, pituitary gland tissue, thyroid gland tissue, adrenal gland tissue, pancreatic tissue, ovarian tissue and testicular tissue.
9. The method of claim 2 , wherein the first motor is a variable speed motor.
10. The method of claim 9 , wherein the first motor drives the impact arm such that the impact arm articulates up and down at a rate of up to 30,000 rpm, making physical contact with the processing container.
11. The method of claim 10 , wherein the first motor drives the up and down articulation of the impact arm at a rate of up to 5,000 rpm.
12. The method of claim 10 , wherein the articulation of the impact arm generates mechanical impacts that are imparted to an outside surface of the processing container that break down the tissue and separate the stromal vascular fraction from the adipose tissue.
13. The method of claim 12 , wherein the first motor controls the speed at which the impact arm makes contact with the processing container.
14. The method of claim 13 , wherein the first motor is controlled by a microprocessor.
15. The method of claim 1 , wherein the processing container is a cartridge.
16. The method of claim 1 , wherein the processing container does not contain enzymes other than those in the tissue.
17. The method of claim 1 , wherein the impacts do not cause mixing of the stromal vascular fraction and the remaining tissue.
18. The method of claim 2 , wherein the processing device comprises a second motor that controls movement of the platform.
19. The method of claim 18 , wherein the second motor controls vertical movement of the platform.
20. The method of claim 4 , wherein the mechanical impacts are imparted to an outside surface of the processing container.