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 stem cell, comprising:
obtaining brown fat tissue;
isolating a stem cell from the brown fat tissue; and
culturing the stem cell in a medium not comprising an animal product, thereby generating a stem cell.
2 . The method of claim 1 , wherein the brown fat tissue is from a sample obtained from a subject.
3 . The method of claim 1 , wherein the stem cell is positive for one or more of the following cell surface markers: CD63, CD90, HLA ABC, CD105, CD73, CD166, CD 9, CD44.
4 . The method of claim 1 , wherein the stem cell is negative for one or more of the following cell surface markers: CD34, CD19, CD86, HLA DR, Lin, CD106, CD80, CD 117.
5 . A method of treating a metabolic disorder in a subject, comprising:
removing a stem cell from a donor;
differentiating the stem cell into a brown adipocyte in a medium not comprising an animal product; and
administering the brown adipocyte to a fat deposit within the subject, thereby treating the metabolic disorder in the subject.
6 . The method of claim 5 , wherein the metabolic disorder is selected from the group consisting of obesity, central obesity, diabetes, hypertension, cardiac deficiency, ischemic cardiac disease, high blood pressure, triglyceride dyslipidemia, HDL dyslipidemia, cholesterol dyslipidemia, elevated fasting plasma glucose, leptin dysregulation, and adipon dysregulation.
7 . The method of claim 5 , wherein the brown adipocyte is administered by subcutaneous injection.
8 . The method of claim 5 , wherein the brown adipocyte is administered by systemic injection.
9 . The method of claim 5 , wherein the medium contains a differentiation agent selected from the group consisting of insulin, dexamethasone, isobutylmethylxanthine, and indomethacin.
10 . A method of treating obesity in a subject, comprising:
removing brown fat from a subject; and
administering the brown fat to a fat deposit within the subject, thereby treating obesity in the subject.
11 . The method of claim 10 , wherein the fat deposit is a white fat deposit.
12 . The method of claim 10 , wherein the fat deposit is a brown fat deposit.
13 . A method of identifying a compound involved in metabolism regulation, comprising:
isolating a stem cell from the brown fat tissue;
culturing the stem cell in a medium not comprising an animal product;
contacting the stem cell with a test compound;
determining the level of metabolic activity of the stem cell in the presence of the test compound; and
comparing the level of metabolic activity of the stem cell in the presence of the test compound to a level of metabolic activity of the stem cell in the absence of the test compound,
wherein a level of metabolic activity in the presence of the test compound that is different from a level of metabolic activity in the absence of the test compound identifies the test compound as a compound involved in metabolism regulation.
14 The method of claim 13 , wherein the determining comprises measuring adipogenesis.
15 . The method of claim 14 , comprising detecting the levels of one or more of PRDM16, BMP7, UCP1, UCP2, or MYF5.
16 . The method of claim 13 , wherein the test compound is selected from the group consisting of protein, antibody, peptide, mutein, polynucleotide, nucleic acid aptamer, and small molecule.
17 . A method of differentiating a stem cell into a brown fat cell, the method comprising:
contacting a stem cell with a medium not comprising an animal product, thereby generating a brown fat stem cell.
18 . The method of claim 17 , wherein the medium comprises an agent selected from the group consisting of: insulin, dexamethasone, isobutylmethylxanthine, indomethacin, T3, rosiglitazone, FNDC5 and combinations thereof
19 . The method of claim 17 , wherein the medium comprises insulin, dexamethasone, isobutylmethylxanthine, and indomethacin.
20 . The method of claim 17 , wherein the brown fat stem cell expresses PRDM-16, PGC-1, or a combination thereof.
21 . The method of claim 17 , wherein the brown fat tissue is from a sample obtained from a subject.