BRIEF DESCRIPTION OF THE DRAWINGS
For the purpose of illustrating the invention, there are depicted in the drawings certain embodiments of the invention. However, the invention is not limited to the precise arrangements and instrumentalities of the embodiments depicted in the drawings.
FIG. 1, comprising FIGS. 1A through 1C, is a series of images depicting adipogenesis in human adipose-derived adult stem cells. FIG. 1A depicts Passage 1 human adipose-derived adult stem cells under control (undifferentiated) or differentiated conditions after staining with Oil Red O at 10× magnification. FIG. 1B depicts the same cells with no magnification. FIG. 1C is a graph illustrating the percentage of Oil Red O surface area staining for an entire plate.
FIG. 2, comprising FIGS. 2A through 2C, is a series of images depicting two-dimensional polyacrylamide gel electrophoresis of protein lysates prepared from human adipose-derived adult stem cells in the undifferentiated (FIG. 2B) and differentiated (FIG. 2C) condition following induction. FIG. 2A is an image of a composite gel prepared based on features conserved on replicate gels prepared from protein extracts obtained from individual donors.
FIG. 3, comprising FIGS. 3A and 3B, is a series of graphs depicting the functionality and subcellular localization of identified proteins in undifferentiated human adipose-derived adult stem cells. The subcellular localization (FIG. 3A) and functionality (FIG. 3B) of the protein categories detailed graphically as percentages are based on n=175 individual proteins identified in the undifferentiated adipocyte cells. Abbreviations: C, Cytoplasm; CaBP, Calcium Binding Protein; Chap, Chaperone; Cytoskel, Cytoskeleton; ECM, Extracellular matrix; ER, Endoplasmic Reticulum; GBP, Guanine nucleoside Binding Protein; M, Mitochondria; Metab, Metabolism; N, Nucleus; NR, Not Reported; ProtDeg, Protein Degradation; ProtProc, Protein Processing; RNABP, RNA Binding Protein. The “Other” Location category includes the Golgi, Lysosome, Plasma Membrane, Ribosome, and Secreted while the “Other” Function category includes Amyloid Binding Protein, Cytokine, Ion Channel, Iron Binding Protein, Signal Transduction, and Transcription.
FIG. 4, comprising FIGS. 4A through 4H, is a series of images depicting protein features from 2-D gels prepared with protein lysates from undifferentiated and differentiated human adipose-derived adult stem cells. FIGS. 4A through 4D represent undifferentiated adipose-derived adult stem cells, FIGS. 4E through 4H represent differentiated adipose-derived adult stem cells. The SSP numbers identify the following protein(s): 3101, Fatty acid binding protein-adipocyte; 7204, Heat shock protein 20-like protein; 3107, Stathmin; 6521, Elfin/PDZ and LIM Domain Protein 1 and LIM and SH3 Domain Protein 1. The arrows indicate the location of the protein features.
FIG. 5, comprising FIGS. 5A and 5B, is a series of images depicting immunoblot analysis of heat shock proteins and chaperones. Protein lysates from the undifferentiated (U) and differentiated (D) adipose-derived adult stem cells obtained from two individual donors were detected using antibodies than bind heat shock proteins and chaperones. The average signal intensity ratio (D/U) of the differentiated to undifferentiated cell lysates is indicated.
FIG. 6 is an image depicting the immunoblot detection of Heat Shock Protein 27 phosphoserine 82. Protein lysates from the undifferentiated (U) and differentiated (D) adipose-derived adult stem cells from four individual donors were detected using antibodies to heat shock protein 27 phosphoserine 82 and all isoforms of heat shock protein 27. The average signal intensity ratio (D/U) of the differentiated to undifferentiated cell lysates is indicated.
FIG. 7 is an image depicting the immunoblot detection of Crystallin alpha phosphoproteins. Protein lysates from the undifferentiated (U) and differentiated (D) adipose-derived adult stem cells from four individual donors were detected using antibodies to crystalline alpha β (heat shock protein beta) phosphoserines 19, 45, and 59. The average signal intensity ratio (D/U) of the differentiated to undifferentiated cell lysates is indicated.
FIG. 8, comprising FIGS. 8A through 8J, is a table depicting the protein features of undifferentiated human adipose-derived adult stem cells.
FIG. 9, comprising FIGS. 9A through 9C, is a table depicting proteins that are upregulated ≧2-fold with adipogenesis in human adipose-derived adult stem cells.
FIG. 10 is a table depicting proteins that are downregulated ≧3-fold with adipogenesis in human adipose-derived adult stem cells.
FIG. 11, comprising FIGS. 11A through 11C, is a table depicting proteins identified in undifferentiated and differentiated human adipose-derived adult stem cells.
FIG. 12, comprising FIGS. 12A through 12D, is a table depicting secreted proteins identified in undifferentiated and differentiated human adipose-derived adult stem cells.
FIG. 13, comprising FIGS. 13A through 13X, is a series of images depicting protein features from two-dimensional gels prepared with protein lysates from undifferentiated and differentiated human adipose-derived adult stem cells. FIGS. 13A through 13H depict undifferentiated, and FIGS. 121 through 13P depict differentiated adipose-derived adult stem cells. The SSP numbers identify the following proteins: 3705, pregnancy zone protein precursor; 3208, adiponectin precursor; 1301, calumenin precursor; 4202, heat shock protein 27 (beta 1); 5301, pigment epithelial derived factor precursor (serpin); 5302, pigment epithelial derived factor; 3203, placental thrombin inhibitor (serpin 6); and 7302, plasminogen activator inhibitor I PAI-1. The arrows indicate the location of the protein features. FIGS. 13P through 13X are a series of bar graphs indicating the relative abundance of the spot on the gels from undifferentiated cells (first four bars) versus differentiated (last four bars) adipose-derived adult stem cells.
FIG. 14, comprising FIGS. 14A through 14C, is a series of images depicting two-dimensional polyacrylamide gel electrophoresis of adipose-derived adult stem cells. FIG. 14B depicts gel electrophoresis results from undifferentiated adipose-derived adult stem cells, FIG. 14C depicts gel electrophoresis results from differentiated adipose-derived adult stem cells, and FIG. 14A depicts a master composite of the gels from the two conditions, prepared based on features conserved on replicate gels prepared from protein extracts obtained from the donors.
FIG. 15, comprising FIGS. 15A through 15E, is a series of graphs depicting quantitative real time PCR results for various secreted proteins from undifferentiated and differentiated adipose-derived adult stem cells. FIG. 15A is a graph depicting quantitative real time PCR results from protease C1 inhibitor normalized to cyclophilin B for undifferentiated and differentiated human adipose-derived adult stem cells from individual donors. Values are the mean±S.D. for triplicate determinations for each donor sample. FIG. 15B is a graph depicting quantitative real time PCR results from plasminogen activator inhibitor-1 (PAI-1) normalized to cyclophilin B for undifferentiated and differentiated human adipose-derived adult stem cells from individual donors. Values are the mean±S.D. for triplicate determinations for each donor sample. FIG. 15C is a graph depicting quantitative real time PCR results from pigmented epithelial derived factor (PEDF) normalized to cyclophilin B for undifferentiated and differentiated human adipose-derived adult stem cells from individual donors. Values are the mean±S.D. for triplicate determinations for each donor sample. FIG. 15D is a graph depicting quantitative real time PCR results from crystallin αB normalized to cyclophilin B for undifferentiated and differentiated human adipose-derived adult stem cells from individual donors. Values are the mean±S.D. for triplicate determinations for each donor sample. FIG. 15E is a graph depicting quantitative real time PCR results from heat shock protein 27 normalized to cyclophilin B for undifferentiated and differentiated human adipose-derived adult stem cells from individual donors. Values are the mean±S.D. for triplicate determinations for each donor sample.
Patent Application
20070207504
