SYSTEMS AND METHODS FOR MANIPULATION OF REGENERATIVE CELLS FROM ADIPOSE TISSUE
The invention provides methods for manipulating regenerative cells from adipose tissue. Specifically, it provides methods for enrichment of desired cells and enhancement of their therapeutic effects.
1 . A method for processing regenerative cells from adipose tissue comprising:
(a) introducing adipose tissue into a tissue collection container of a device configured to harvest regenerative cells from adipose tissue, while maintaining a closed, sterile fluid/tissue pathway;
(b) reducing the presence of free lipids and peripheral blood elements from said adipose tissue in the tissue collection container;
(c) disaggregating said adipose tissue in the tissue collection container to generate a cell suspension;
(d) separating the regenerative cells from the acellular component in the suspension;
(e) concentrating the regenerative cells; and
(f) manipulating the regenerative cells to obtain processed regenerative cells, wherein the manipulation is selected from the group consisting of exposure to hypoxic conditions, exposure to hyperoxic conditions, exposure to UV light, exposure to infared light, exposure to ultrasonic stimulation, and exposure to electrical stimulation.
2 . The method of claim 1 , further comprising delivering the processed regenerative cells to a patient.
3 . The method of claim 1 , further comprising heating or cooling the regenerative cells.
4 . The method of claim 1 , wherein the separating comprises density gradient centrifugation, or continuous flow centrifugation, or both.
5 . The method of claim 1 , wherein the separating comprises adhering sample components to a solid phase surface.
6 . The method of claim, wherein the solid phase surface is selected from the group consisting of tissue culture plastic, plastic beads, glass beads, and scaffolds or any combination thereof.
7 . The method of claim 1 , further comprising exposing the regenerative cells to an additive.
8 . The method of claim 7 , wherein the additive is selected from the group consisting of a tissue fragment, a growth factor, a cell differentiation factor, an immunosuppressive agent, an anti-apoptotic agent, and an anti-inflammatory agent.
9 . The method of claim 1 , further comprising combining the processed regenerative cells with a biologically compatible scaffold or carrier.
10 . The method of claim 1 , further comprising contacting the regenerative cells with DNAse I.
11 . The method of claim 1 , wherein the disaggregating comprises mechanical disaggregation of the adipose tissue.
12 . The method of claim 11 wherein the mechanical disaggregation comprises ultrasonic disaggregation.
13 . The method of claim 1 , wherein the separating comprises filtration.
14 . The method of claim 1 , wherein the manipulation comprises exposure to hypoxic conditions comprising about 1% O 2 .
15 . The method of claim 1 , wherein the regenerative cells comprise adipose-derived stem cells and endothelial progenitor cells.
16 . The method of claim 9 , wherein the biologically compatible scaffold or carrier is a resorbable scaffold.
17 . The method of claim 9 , wherein the biologically compatible scaffold or carrier is selected from the group consisting of a hyaluronon-based scaffold, an apatite coated scaffold, a hydrogel, and a collagen gel.
18 . The method of claim 1 , further comprising formulating the processed regenerative cells for injection.
19 . The method of claim 1 , wherein the degree of disaggregation is determined.
20 . The method of claim 19 , wherein the degree of disaggregation is determined by measuring current flow through the cell suspension, the optical density of the cell suspension, a color change in the cell suspension, or a color change in a waste solution generated during processing.