NON-NATURALLY OCCURING THREE-DIMENSIONAL (3D) BROWN ADIPOSE-DERIVED STEM CELL AGGREGATES, AND METHODS OF GENERATING AND USING THE SAME
The present application provides non-naturally occurring 3D brown adipose-derived stem cell (BADSC) aggregates, methods of making the 3D BADSC aggregates, and methods of using the 3D BADSC aggregates.
1 . A non-naturally occurring three-dimensional brown adipose derived stem cell aggregate wherein the three-dimensional brown adipose derived stem cell aggregate comprises brown adipose-derived stem cells that express one or more brown adipocyte gene in the absence of differentiation medium.
2 . The non-naturally occurring three-dimensional brown adipose derived stem cell aggregate of claim 1 , wherein the one or more brown adipocyte gene is selected from a group consisting of PPARα, PPARγ, PGC1β, PRDM16, CEBPD, CEBPB, CEBPA, TFAM, PGC1α, and PGC1β.
3 . The non-naturally occurring three-dimensional brown adipose derived stem cell aggregate of claim 1 , wherein the aggregate forms in a non-adherent environment.
4 . The non-naturally occurring three-dimensional brown adipose derived stem cell aggregate of claim 1 , wherein aggregate produces extracellular biologics selected from a group consisting of exosomes, microRNA, cytokines, proteins, and adipokines.
5 . An encapsulation system comprising the non-naturally occurring three-dimensional brown adipose derived stem cell aggregate of claim 1 .
6 . The encapsulation system of claim 5 , wherein the encapsulation system is selected from the group consisting of alginate microcapsules, cellulose hydrogels, red blood cells, porous polymer membranes, 3D biological scaffolds, polymers, PEG-based hydrogels, non-hydrogel beads, and matrigel.
7 . The encapsulation system of claim 5 , wherein the encapsulation system is an encapsulation medical device.
8 . A method of making a non-naturally occurring three-dimensional brown adipose derived stem cell aggregate, the method comprising:
loading brown adipose derived stem cells grown in a two-dimensional (2D) culture into a non-adherent culture plate; and
centrifuging the non-adherent culture plate to uniformly position the brown adipose-derived stem cells in the non-adherent culture plate, thereby forming three-dimensional brown adipose derived stem cell aggregates.
9 . The method of claim 8 , further comprising:
prior to the loading, culturing the brown adipose derived stem cells in a two-dimensional (2D) culture using growth medium under normoxia or hypoxia.
10 . A method of making a three-dimensional brown adipose tissue in an encapsulation system, the method comprising:
forming non-naturally occurring three-dimensional brown adipose derived stem cell aggregates;
loading the non-naturally occurring three-dimensional brown adipose derived stem cell aggregates into the encapsulation system;
differentiating the non-naturally occurring three-dimensional brown adipose derived stem cell aggregates into brown adipose tissue in a first differentiation medium; and
differentiating the non-naturally occurring three-dimensional brown adipose derived stem cell aggregates into brown adipose tissue in a second differentiation medium.
11 . The method of claim 10 , wherein the encapsulation system is selected from the group consisting of alginate microcapsules, cellulose hydrogels, red blood cells, porous polymer membranes, 3D biological scaffolds, polymers, PEG-based hydrogels, non-hydrogel beads, and matrigel.
12 . The method of claim 10 , wherein the encapsulation system is an encapsulation medical device.
13 . The method of claim 10 , wherein the first differentiation medium comprises dexamethasone, IBMX, and T3.
14 . The method of claim 10 , wherein the second differentiation medium comprises T3 and rosiglitazone.
15 . A method of treating a patient with a disorder, the method comprising:
forming non-naturally occurring three-dimensional brown adipose derived stem cell aggregates;
loading the non-naturally occurring three-dimensional brown adipose derived stem cell aggregates into an encapsulation system;
differentiating the non-naturally occurring three-dimensional brown adipose derived stem cell aggregates into brown adipose tissue in a first differentiation medium;
differentiating the non-naturally occurring three-dimensional brown adipose derived stem cell aggregates into brown adipose tissue in a second differentiation medium; and
delivering the brown adipose tissue to the patient with the disorder.
16 . The method of claim 15 , wherein the encapsulation system is selected from the group consisting of alginate microcapsules, cellulose hydrogels, red blood cells, porous polymer membranes, 3D biological scaffolds, polymers, PEG-based hydrogels, non-hydrogel beads, and matrigel.
17 . The method of claim 15 , wherein the encapsulation system is an encapsulation medical device.
18 . The method of claim 15 , wherein the first differentiation medium comprises dexamethasone, IBMX, and T3.
19 . The method of claim 15 , wherein the second differentiation medium comprises T3 and rosiglitazone.
20 . The method of claim 15 , wherein the disorder is a metabolic disorder, an endocrine disorder, a cardiovascular disorder, or a liver disease.
21 . The method of claim 20 , wherein the metabolic disorder is obesity or diabetes.