Multiple Mesodermal Lineage Differentiation Potentials for Adipose Tissue-Derived Stromal Cells and Uses Thereof
AssigneeArtecel Sciences, Inc.
InventorsJeffrey Martin Gimble, Yuan-Di Chang Halvorsen, William O. Wilkison
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Abstract
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.
Claims18
A method for differentiating adipose tissue derived stromal cells into hematopoietic supporting stromal cells which will proliferate and differentiate along the myeloid lineage pathway or the B-lineage lymphoid pathway, comprising:
- a) plating said stromal cells at a density of about 30,000 cells per cm2 in chamber slides;
- b) maintaining cells in culture for about 8 days in a medium containing Dulbecco's Modified Eagle's Medium (DMEM) or Ham's F-10;
- c) supplementing said medium with:
- (i) 1 to 20% fetal bovine serum (ii) an antibiotic (iii) interleukins (iv) stem cell factor (v) flt-3 ligand (vi) macrophage-colony stimulating factor (vii) granulocyte-monocyte colony stimulating factor (viii) erythropoetin (ix) thrombopoietin (x) osteoprotegerin ligand (xi) dexamethasone (xii) hydrocortisone (xiii) 1,25 dihydroxy vitamin D3 and (xiv) 2-mercaptoethanol;
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- d) examining the expression of cell surface proteins which are consistent with cells of the myeloid lineage or B-lymphoid lineage using a variety of techniques which include but are not limited to immunohistochemistry, flow cytometry, immunofluorescence and mRNA expression in cell populations.
Dependencies are inferred from the claim text.