Multiple mesodermal lineage differentiation potentials for adipose tissue-derived stromal cells and uses thereof
View Patent ↗The invention relates to methods and compositions for the differentiation of stromal cells from adipose tissue into hematopoietic supporting stromal cells and myocytes of both the skeletal and smooth muscle type. The cells produced by the methods are useful in providing a source of fully differentiated and functional cells for research, transplantation and development of tissue engineering products for the treatment of human diseases and traumatic tissue injury repair.
1. A medium for differentiating adipose tissue derived stromal cells into hematopoietic supporting stromal cells comprising a chemically defined medium having or supplemented with the following components: (i) 0% to 20% fetal bovine serum; (ii) antibiotic; (iii) interleukins; (iv) about 5 pg/ml to 1 ng/ml stem cell factor; (v) about 5 pg/ml to 1 ng/ml flt-3ligand; (vi) about 5 pg/ml to 1 ng/ml macrophage-colony stimulating factor; (vii) about 5 pg/ml to 1 ng/ml granulocyte-monocyte colony stimulating factor; (viii) about 5 units/ml to 1000 units/ml erythropoietin; (ix) about 5 pg/ml to 1 ng/ml thrombopoietin; (x) about 5 pg/ml to 1 ng/ml osteoprotegerin ligand; (xi) about 1 nM to 100 nM dexamethasone; (xii) about 1 nM to 100 nM hydrocortisone; (xiii) about 1 nM to 100 nM 1,25 dihydroxy vitamin D 3 ; and (xiv) about 10 uM to 100 uM 2-mercaptoethanol and wherein components (ii)-(xiv) are present.
2. The medium of claim 1 , wherein said chemically defined medium is selected from the group consisting of: DMEM, aMEM, and RPMI media 1640.
3. The medium of claim 1 , wherein said antibiotic is penicillin.
4. The medium of claim 1 , wherein said antibiotic is streptomycin.
5. The medium of claim 3 , wherein said penicillin is present in amounts from about 10 units per ml to 200 units per ml in the total volume of the medium.
6. The medium of claim 4 , wherein said streptomycin is present in amounts from about 10 pg per ml to 200 pg per ml in the total volume of the medium.
7. The medium of claim 1 , wherein said interleukins are selected from the group consisting of: interleukin-1, interleukin-3, interleukin-6, interleukin-7, interleukin 11 and interleukin-12.
8. The medium of claim 7 , wherein the total amount of all interleukins is from about 5 pg/ml to 1 ng/ml.
9. The medium of claim 1 , wherein said medium is at a temperature of about 33° C. for myeloid cells.
10. The medium of claim 1 , wherein said medium is at a temperature of about 37° C. for B-lineage lymphoid cells.