Brown fat cell compositions and methods
Methods of developing and using cell lines, such as stem cell lines, for therapeutic or cosmetic use. In one embodiment, the cell lines are used to treat a wide range of degenerative and metabolic disorders including, but not limited to, obesity, diabetes, hypertension, and cardiac deficiency. Also described are methods of using such cell lines to screen for compounds that play a role in regulating a variety of processes.
1. A method of generating a population of human brown fat adipocytes, the method comprising:
removing a human stem cell from human brown adipose tissue;
differentiating the human stem cell into a population of human brown fat adipocytes in a culture medium under a selective pressure of 0 to 5% oxygen, wherein the culture medium does not comprise a xenogenic animal product and comprises fibronectin type III domain-containing protein 5 (FNDC), insulin, dexamethasone, isobutylmethylxanthine, indomethacin, triiodothyronine (T3), rosiglitazone, and 0.5% to 20% human platelet lysate.
2. The method of claim 1 , further comprising: providing the population of human brown fat adipocytes for implantation into a subject.
3. The method of claim 1 , wherein the selective pressure is 1 to 5% oxygen.
4. The method of claim 2 , wherein the population of human brown fat adipocytes is allogeneic.
5. The method of claim 2 , wherein the population of human brown fat adipocytes is autogeneic.
6. The method of claim 1 , wherein the culture medium comprises 10-20% human platelet lysate.
7. The method of claim 4 , further comprising:
implanting the allogeneic population of human brown fat adipocytes into a subject to be treated.
8. The method of claim 7 , wherein the allogenic population of human brown fat adipocytes comprises 10 5 -10 10 .
9. The method of claim 5 , further comprising:
implanting the autogeneic population of human brown fat adipocytes into a subject to be treated.
10. The method of claim 9 , wherein the autogeneic population of human brown fat adipocytes comprises 10 5 -10 10 cells.
11. The method of claim 1 , further comprising:
isolating exosomes from the culture medium of the population of human brown fat adipocytes.
12. A method of generating a population of human brown fat adipocytes, the method comprising:
differentiating a human stem cell into a population of human brown fat adipocytes in a first culture medium under a selective pressure of 0 to about 5% oxygen, wherein the first culture medium does not comprise a xenogenic animal product and consists of: insulin, dexamethasone, isobutylmethylxanthine, indomethacin, triiodothyronine (T3), rosiglitazone, and 0.5 to 20% human platelet lysate;
removing the first culture medium;
further differentiating the human stem cell into the population of human brown fat adipocytes in a second culture medium under a selective pressure of 0 to about 5% oxygen wherein the second culture medium does not comprise a xenogenic animal product and consists of: insulin, dexamethasone, isobutylmethylxanthine, indomethacin, triiodothyronine (T3), rosiglitazone, fibronectin type III domain-containing protein 5 (FNDC5), and 0.5 to 20% human platelet lysate
wherein the human stem cell is removed from human brown adipose tissue.