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 tissue processing device, comprising:
a processing container for containing raw adipose tissue;
a platform onto which the processing container is attached;
an impact arm; and
a variable speed motor that drives the impact arm such t hat the impact arm articulates up and down at a rate of up to 30,000 rpm, making physical contact with the processing container;
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 raw adipose tissue and separate stem cells from the adipose tissue; and
the motor controls the speed at which the impact arm makes contact with the processing container; and
wherein the motor comprises gearing capable of reducing the speed of the impact arm.
2. The device of claim 1 , wherein the motor drives the up and down articulation of the impact arm at a rate selected from the group consisting of: up to 20,000 rpm, up to 10,000 rpm, up to 5,000 rpm, and between 3,000 and 30,000 rpm.
3. The device of claim 1 , wherein the processing container is a bag.
4. The device of claim 3 , wherein the bag comprises an outlet in fluid connection with the interior of the container.
5. The device of claim 3 , wherein the bag comprises more than one outlet in fluid connection with the interior of the container.
6. The device of claim 4 , wherein the bag further comprises an inlet in fluid connection with the interior of the container.
7. The device of claim 1 , wherein the processing container does not contain enzymes other than those in the tissue.
8. The device of claim 1 , wherein the impacts do not cause mixing of the stem cells and the remaining tissue.
9. The device of claim 1 , wherein the motor is controlled by a microprocessor.
10. A method of isolating stem cells from adipose tissue, comprising:
placing an adipose tissue sample from a subject into the processing container of the device of claim 1 ;
subjecting the tissue sample to force from mechanical impacts to break down the tissue; and
separating a stem cell fraction from the adipose tissue;
wherein:
the impact arm does not physically contact the adipose tissue.
11. The method of claim 10 , wherein the adipose tissue is washed one or more times prior to being subjected to force from the mechanical impacts.
12. The method of claim 10 , wherein the force from the mechanical impacts is delivered to the adipose tissue through a wall of the processing container.
13. The method of claim 10 , wherein the separating comprises allowing the adipose tissue to separate from an aqueous layer, the aqueous layer comprising the stem cells.
14. The method of claim 10 , wherein the stem cell fraction is separated from the adipose tissue in 30 minutes or less.