Systems and methods for processing cells
The present invention efficiently and cost-effectively extracts and collects cells from a tissue. The inventors have discovered that the tissue can be effectively fragmented and the resulting cells can be purified using a system or kit with multiple components. An advantage of the present invention is that tissue processing takes place in a closed system such that sterility can be maintained throughout the process, even if certain components are removed during processing, for example through the use of valves, clamps, and heat seals. Furthermore, any or all of the steps can be automated or manually accomplished, according to the specific needs of the application or the user.
1. A tissue mincing tool comprising:
a compartment for a tissue sample;
a solid member actuatable by a handle and moveable within the compartment;
a rotatable cutting surface at one end of the compartment; and
a sterile, sealed container,
wherein the compartment is arranged to position the tissue sample between the solid member and the rotatable cutting surface and actuation of the handle in one direction causes the solid member to move only towards the rotatable cutting surface,
and further wherein the rotatable cutting surface separates the compartment from the sterile, sealed container, such that a tissue sample that is pressed, via the solid member, into contact with and through the rotatable cutting surface is deposited as a minced tissue within the sterile, sealed container.
2. The tissue mincing tool of claim 1 , wherein the rotatable cutting surface is dimensioned to mince the tissue sample into fragments having an average cross-section no greater than four square millimeters.
3. The tissue mincing tool of claim 1 , wherein the rotatable cutting surface is dimensioned to mince the tissue sample into fragments, and further comprising a second cutting surface for reducing an average cross-section of the fragments.
4. The tissue mincing tool of claim 1 , further comprising at least one mincer screen positioned in proximity to the rotatable cutting surface.
5. The tissue mincing tool of claim 4 , wherein the at least one mincer screen is moveable towards the tissue sample.
6. The tissue mincing tool of claim 1 , further comprising a suction cup for stabilizing the tissue mincing tool during operation thereof.
7. The tissue mincing tool of claim 1 , wherein the sterile, sealed container comprises at least one sealed access port permitting the sterile introduction of a fluid into the container.
8. The tissue mincing tool of claim 1 , wherein an interior surface of the compartment is threaded.
9. The tissue mincing tool of claim 1 , further comprising a gasket for sealing the compartment.
10. The tissue mincing tool of claim 1 , wherein a portion of the compartment near its end has a substantially constant cross-section.
11. The tissue mincing tool of claim 10 , wherein the solid member is shaped to mate with and to fill the portion of the compartment.
12. The tissue mincing tool of claim 1 , wherein the rotatable cutting surface is moveable towards the tissue sample.
13. The tissue mincing tool of claim 1 , further comprising a shaft crank for moving the rotatable cutting surface towards the tissue sample.
14. A method of mincing a tissue sample positioned between the solid member and the rotatable cutting surface of the tissue mincing tool of claim 1 , the method comprising:
pressing the tissue sample, via the solid member, into contact with and through the rotatable cutting surface of the tissue mincing tool.
15. A method of digesting a tissue sample, the method comprising:
mincing the tissue sample according to the method of claim 14 and, optionally, transferring the minced tissue to a second sterile, sealed container; and
injecting an enzyme, whereby the enzyme digests the minced tissue.
16. The method of claim 15 , wherein the enzyme is a collagenase.
17. A method of separating cells from a tissue sample, the method comprising:
mincing the tissue sample according to the method of claim 14 ; and
removing undigested tissue.
18. The method of claim 17 , further comprising diluting the tissue sample, wherein the undigested tissue is subsequently removed by filtering.
19. The method of claim 14 , wherein the tissue sample comprises Wharton's Jelly stem cells.
20. The method of claim 18 , further comprising sedimenting a filtrate generated by the filtering.
21. The method of claim 20 , further comprising re-suspending sedimented cells and filtering the re-suspended sedimented cells.
22. The method of claim 19 , wherein the tissue sample is substantially free of blood vessels.
23. The tissue mincing tool of claim 1 , wherein the rotatable cutting surface comprises an automated cutting system.