Methods related to minimally polarized functional units
Provided herein are constructs of micro-aggregate multicellular, minimally polarized grafts containing Leucine-rich repeat-containing G-protein coupled Receptor (LGR) expressing cells for wound therapy applications, tissue engineering, cell therapy applications, regenerative medicine applications, medical/therapeutic applications, tissue healing applications, immune therapy applications, and tissue transplant therapy applications which preferably are associated with a delivery vector/substrate/support/scaffold for direct application.
1. A method, comprising:
a) separating fat and hypodermal elements from a mammalian tissue specimen ex vivo to provide remaining cutaneous elements containing an epidermal compartment, a dermal compartment, and a follicular compartment;
b) cutting through the epidermal compartment, the dermal compartment, and the follicular compartment to open the follicular compartment and prepare a cellular micro-aggregate comprising an epidermal segment, a dermal segment, and a segment of the follicular compartment, wherein the segments are interconnected and wherein the segment of the follicular compartment comprises living LGR-expressing stem cells that are exposed;
c) processing the fat and hypodermal elements to provide a mesenchymal-derived cellular population; and
d) adding the mesenchymal-derived cellular population to the cellular micro-aggregate to provide a composition, wherein the composition is capable of regenerating functional tissue comprising epidermis, dermis, and appendages when transplanted to a damaged tissue site in a subject in need thereof.
2. The method of claim 1 , wherein the living LGR-expressing stem cells comprise LGR4-expressing stem cells, LGR5-expressing stem cells, LGR6-expressing stem cells, or any combination thereof.
3. The method of claim 1 , further comprising adding a cornification medium to the composition.
4. The method of claim 1 , further comprising cryopreserving the composition.
5. The method of claim 1 , wherein the mesenchymal-derived cellular population comprises an adipocyte population, a stromal vascular fraction, or any combination thereof.
6. The method of claim 1 , further comprising adding the composition to a delivery substrate selected from scaffolding, matrix, particle, cells, fiber, or any combination thereof.
7. The method of claim 1 , further comprising adding enhancing factors to the composition.
8. The method of claim 1 , wherein b) comprises cutting through the epidermal compartment, the dermal compartment, and a bulge of the follicular compartment.
9. The method of claim 1 , wherein b) comprises segmenting the epidermal compartment, the dermal compartment, and a bulb of the follicular compartment.
10. The method of claim 1 , further comprising adding the composition to a pharmaceutically acceptable carrier.
11. A method, comprising applying the composition produced by the method of claim 1 to a select target.
12. The method of claim 11 , wherein the select target is selected from a tissue region, a wound, a void, a defective tissue, or any combination thereof.
13. The method of claim 11 , wherein the select target is an injury involving single or multiple tissue elements.
14. The method of claim 11 , wherein the applying is direct or indirect.
15. The method of claim 11 , wherein the applying is direct and a delivery technique is selected from transplantation, implantation, directed seeding, directed migration, directed tracking, in setting, laminating, injection, or combinations thereof.
16. A method comprising:
a) separating fat and hypodermal elements from a mammalian tissue specimen ex vivo to provide remaining cutaneous elements containing an epidermal compartment, a dermal compartment, and a follicular compartment;
b) segmenting epidermal compartment, the dermal compartment, and the follicular compartment to open the follicular compartment and prepare an epidermal segment, a dermal segment, and a segment of the follicular compartment, wherein the segments are interconnected and wherein the segment of the follicular compartment comprises living LGR-expressing stem cells that are exposed;
c) processing the fat and hypodermal elements to provide a mesenchymal-derived cellular population; and
d) adding the mesenchymal-derived cellular population to the interconnected segments to provide a composition, wherein the composition is capable of regenerating functional tissue comprising epidermis, dermis, and appendages when transplanted to a damaged tissue site in a subject in need thereof.
17. The method of claim 16 , wherein the living LGR-expressing stem cells comprise LGR4-expressing stem cells, LGR5-expressing stem cells, LGR6-expressing stem cells, or any combination thereof.
18. The method of claim 16 , further comprising adding a cornification medium to the composition.
19. The method of claim 16 , further comprising cryopreserving the composition.
20. The method of claim 16 , wherein the mesenchymal-derived cellular population comprises an adipocyte population, a stromal vascular fraction, or any combination thereof.
21. The method of claim 16 , further comprising adding the composition to a delivery substrate selected from scaffolding, matrix, particle, cells, fiber, or any combination thereof.
22. The method of claim 16 , further comprising adding enhancing factors to the composition.
23. The method of claim 16 , further comprising adding the composition to a pharmaceutically acceptable carrier.
24. The method of claim 16 , wherein b) comprises cutting through the epidermal compartment, the dermal compartment, and a bulge of the follicular compartment.
25. The method of claim 16 , wherein b) comprises segmenting the epidermal compartment, the dermal compartment, and a bulb of the follicular compartment.
26. A method, comprising applying the composition produced by the method of claim 16 to a select target.
27. The method of claim 26 , wherein the select target is selected from a tissue region, a wound, a void, a defective tissue, or any combination thereof.
28. The method of claim 26 , wherein the select target is an injury involving single or multiple tissue elements.
29. The method of claim 26 , wherein the applying is direct or indirect.
30. The method of claim 26 , wherein the applying is direct and a delivery technique is selected from transplantation, implantation, directed seeding, directed migration, directed tracking, in setting, laminating, injection, or combinations thereof.