Journal: Aging Cell

Article Title: Stem cell secretome treatment improves whole‐body metabolism, reduces adiposity, and promotes skeletal muscle function in aged mice

doi: 10.1111/acel.14144

Figure Lengend Snippet: Whole‐Body Metabolic Profile. Oxygen consumption (VO 2 – ml/kg/hr) and carbon dioxide production (VCO 2 – ml/kg/hr) across 24 h in comprehensive lab animal monitoring system (CLAMS) for control and secretome‐treated mice (a, b). 24‐h average VO 2 (c) and VCO 2 (d), 12‐h average dark cycle respiratory exchange ratio (RER – VCO 2 /VO 2 ; e), 24‐h average energy expenditure (f; kcal/kg/hr), activity (g; movement – X + Y + Z), and food intake (g) (h). All data presented as mean ± SD, white bars represent controls while blue squares represent secretome‐treated mice. n = 6 for both groups. * indicates significant difference between conditions for the indicated timepoints with * = p < 0.05, ** = p < 0.01, and *** = p < 0.001.

Article Snippet: Cultured media collected from these cells was pooled, sterile filtered, concentrated, and prepared as a USP‐grade cell‐free stem cell‐based secretome product (IMMUNA; Immunis, Inc., Irvine, CA).

Techniques: Control, Activity Assay

Journal: Aging Cell

Article Title: Stem cell secretome treatment improves whole‐body metabolism, reduces adiposity, and promotes skeletal muscle function in aged mice

doi: 10.1111/acel.14144

Figure Lengend Snippet: Whole body Tissue and Physical Function Changes. Weekly changes lean mass (a), fat mass (b), body fat % (c), and body mass (d) for control and secretome‐treated mice. Weekly whole‐body grip strength (e) and change (Δ) in rotarod performance time from baseline (f). Quadriceps mass (g) and fat mass (h) following 4‐weeks. Control mice are represented by black circles and bars while secretome‐treated mice are noted by blue squares and bars. All data presented as mean ± SEM. (a–h) control mice ( n = 15), secretome‐treated mice ( n = 16). Weekly changes analyzed via mixed‐effects model with Holm‐Bonferroni multiple comparisons plus planned comparison t ‐tests at the 4‐week timepoint. # indicates significant difference ( p < 0.05) from baseline for respective group. * indicates significant difference ( p < 0.05) between groups at the indicated time point.

Article Snippet: Cultured media collected from these cells was pooled, sterile filtered, concentrated, and prepared as a USP‐grade cell‐free stem cell‐based secretome product (IMMUNA; Immunis, Inc., Irvine, CA).

Techniques: Control, Comparison

Journal: Aging Cell

Article Title: Stem cell secretome treatment improves whole‐body metabolism, reduces adiposity, and promotes skeletal muscle function in aged mice

doi: 10.1111/acel.14144

Figure Lengend Snippet: Skeletal Muscle Morphology. Average quadriceps fiber cross sectional area μm 2 (a), minimum feret diameter μm 2 (b), and fiber type proportion (c; MyHC – IIa/IIb %) for control ( n = 8) and secretome treated ( n = 8) mice. Size distribution (%) for total (e), IIa (f) and IIb (g) fiber types across 500 μm 2 increments. Representative histochemical image of fiber type including cell border with laminin in blue, MyHC IIa in green, MyHC IIb in red, and scale bar of 50 μm (d). All data presented as mean ± SD, white circles and bars represent controls while blue squares and bars represent secretome‐treated mice. n = 8 for each group. Analyzed via t ‐tests and mixed effects models with Holm‐Bonferroni comparisons. * indicates significant difference between groups at indicated category with * = p < 0.05, ** = p < 0.01, *** = p < 0.001.

Article Snippet: Cultured media collected from these cells was pooled, sterile filtered, concentrated, and prepared as a USP‐grade cell‐free stem cell‐based secretome product (IMMUNA; Immunis, Inc., Irvine, CA).

Techniques: Control

Journal: Aging Cell

Article Title: Stem cell secretome treatment improves whole‐body metabolism, reduces adiposity, and promotes skeletal muscle function in aged mice

doi: 10.1111/acel.14144

Figure Lengend Snippet: Skeletal Muscle Remodeling. Muscle satellite cell content (Pax7 + /DAPI + − co‐localization) corrected to number of fibers (a). Representative image of satellite cell localization with PAX7 + in off‐red/pink, laminin fiber borders in green, DAPI in blue, satellite cells indicate by white arrows, and scale bar of 50 μm 2 (b). Fiber capillarization (CD31 + ) corrected to number of muscle fibers (c). Representative image of capillarization with CD31 + in red, laminin in green, and DAPI in blue (d). Ratio of B‐CHP to COL‐IV (e) and representative image with B‐CHP in purple and COL‐IV in yellow (f). All data presented as mean ± SD, white circles and bars represent controls while blue squares and bars represent secretome‐treated mice. Control mice ( n = 8), secretome‐treated mice ( n = 7–8), right (injected) quadriceps assessed. Analyzed via t ‐tests. * indicates significant difference between conditions for the indicated timepoints with * = p < 0.05, ** = p < 0.01.

Article Snippet: Cultured media collected from these cells was pooled, sterile filtered, concentrated, and prepared as a USP‐grade cell‐free stem cell‐based secretome product (IMMUNA; Immunis, Inc., Irvine, CA).

Techniques: Control, Injection

Journal: Aging Cell

Article Title: Stem cell secretome treatment improves whole‐body metabolism, reduces adiposity, and promotes skeletal muscle function in aged mice

doi: 10.1111/acel.14144

Figure Lengend Snippet: Adipose Morphology and Muscle Lipid Content. Average cellular diameter μm of I‐WAT (a) and E‐WAT (b) depots. Size distribution (%) of I‐WAT (c) and E‐WAT (d) cells across 5 μm increments. Average liver lipid droplet (e) and fibrosis (f; trichrome staining) area (%) as assessed with H&E. Representative image of I‐WAT and E‐WAT depots as well as liver H&E and trichrome staining with scale bar 50 μm (g). Protein phosphorylation status for protein kinase B (Akt) and hormone sensitive lipase (HSL) for I‐WAT and E‐WAT depots and representative western blot image (h). Muscle lipid content including total triglycerides (TAGs) diglycerides (DAGs), ceramides (Cer), and C18:0 ceramide (C18:0 Cer). All data presented as mean ± SD, white circles and bars represent controls while blue squares and bars represent secretome treated groups. (a–g) n = 8 for each group, (h–i) n = 7 for control, n = 6 for secretome, (i) n = 6 for control, n = 8 for secretome. Analyzed via t ‐tests (a, b, e, f, h, i) or two‐way ANOVA with Holm‐Bonferroni comparison (c, d). * indicates significant difference between groups at indicated category with * = p < 0.05, ** = p < 0.01.

Article Snippet: Cultured media collected from these cells was pooled, sterile filtered, concentrated, and prepared as a USP‐grade cell‐free stem cell‐based secretome product (IMMUNA; Immunis, Inc., Irvine, CA).

Techniques: Staining, Phospho-proteomics, Western Blot, Control, Comparison

Journal: Aging Cell

Article Title: Stem cell secretome treatment improves whole‐body metabolism, reduces adiposity, and promotes skeletal muscle function in aged mice

doi: 10.1111/acel.14144

Figure Lengend Snippet: Direct and Indirect Cellular Experiments. Cell culture design for media replacement and cultured media (CM) experiments (a). Average myotube area (%) (b) and myonuclear fusion index ( au ) (c) for control, secretome treated, control CM, and secretome CM conditions in differentiated C2C12 myotubes. Interleukin 6 (IL‐6) content in the culture media collected from secretome treated (4%) and control C2C12 myotubes (Fold change) (d). Average lipid droplet area corrected to DAPI area (Fold Change) in 3T3‐L1 adipocytes for control, 5 and 20% secretome product media replacement (e). Phosphorylated corrected to total Akt protein (fold change) for 20% secretome treated and control 3T3‐L1 adipocytes following overnight fast and insulin (100 nM) stimulation (f). Average lipid droplet area corrected to DAPI area (Fold Change) in 3T3‐L1 adipocytes for control and 20% media replacement with culture media from control and secretome treated C2C12 cells (g). Representative images of myotubes (h) and adipocytes treated with secretome (i) and culture media (j). (b–g) n = 4–7 per group or replicate. Analyzed via one‐way ANOVA with Holm‐Bonferroni comparison (b–c, e, g) or t ‐tests (d, f). * indicates significant difference between groups as indicated with * = p < 0.05, ** = p < 0.01, *** = p < 0.001.

Article Snippet: Cultured media collected from these cells was pooled, sterile filtered, concentrated, and prepared as a USP‐grade cell‐free stem cell‐based secretome product (IMMUNA; Immunis, Inc., Irvine, CA).

Techniques: Cell Culture, Control, Comparison

Journal:

Article Title: The Mouse Secretome: Functional Classification of the Proteins Secreted Into the Extracellular Environment

doi: 10.1101/gr.983703

Figure Lengend Snippet: Origin of Proteins Within the Mouse Secretome

Article Snippet: Secretome protein sequences without an InterPro or SCOP hit , 392.

Techniques: Derivative Assay

Journal:

Article Title: The Mouse Secretome: Functional Classification of the Proteins Secreted Into the Extracellular Environment

doi: 10.1101/gr.983703

Figure Lengend Snippet: Size distribution of the proteins predicted to contain endoplasmic reticulum signal peptides within the PROCREST predicted ORFs of the RIKEN 60,770 FANTOM2 cDNA clone set and the mouse secretome. The total numbers of proteins within 50 amino acid blocks are plotted.

Article Snippet: Secretome protein sequences without an InterPro or SCOP hit , 392.

Techniques:

Journal:

Article Title: The Mouse Secretome: Functional Classification of the Proteins Secreted Into the Extracellular Environment

doi: 10.1101/gr.983703

Figure Lengend Snippet: Origin of RPS Proteins Less Than 100 Amino Acids Within the Mouse Secretome

Article Snippet: Secretome protein sequences without an InterPro or SCOP hit , 392.

Techniques:

Journal:

Article Title: The Mouse Secretome: Functional Classification of the Proteins Secreted Into the Extracellular Environment

doi: 10.1101/gr.983703

Figure Lengend Snippet: Distribution of InterPro domains and SCOP superfamilies associated with the mouse secretome. The proportion of individual InterPro domains (A) or SCOP superfamilies (B) within the mouse secretome relative to the total number of predicted domains within the RPS is plotted as a percentage ratio (Secretome/RPS). One hundred percent represents a domain that is only contained in proteins within the mouse secretome. All RPS protein sequences that could not be classified by the membrane organization methods were excluded (Kanapin et al. 2003).

Article Snippet: Secretome protein sequences without an InterPro or SCOP hit , 392.

Techniques: Membrane

Journal:

Article Title: The Mouse Secretome: Functional Classification of the Proteins Secreted Into the Extracellular Environment

doi: 10.1101/gr.983703

Figure Lengend Snippet: Gene Ontology classification of the proteins within the mouse secretome.

Article Snippet: Secretome protein sequences without an InterPro or SCOP hit , 392.

Techniques:

Journal:

Article Title: The Mouse Secretome: Functional Classification of the Proteins Secreted Into the Extracellular Environment

doi: 10.1101/gr.983703

Figure Lengend Snippet: Hierarchical clustering of gene expression data for 970 FANTOM2 cDNAs encoding secretome proteins across 22 mouse tissues. Results are presented in a dendrogram displaying related groups of genes (on the Y-axis) and tissues (on the X-axis). The relative expression of each cDNA is represented as a log2 ratio of hybridization signals between each tissue and a common reference mRNA sample (total 17.5 dpc mouse embryo). Blocks of red signal indicate high levels of expression in the tissue relative to the reference RNA. Blocks of green signify low levels of expression relative to the reference RNA. Grey blocks indicate data points that are missing. Along the right-hand side of the dendrogram are several yellow boxes labeled A to D, which define the location of four tissue-restricted clusters. (A) Neural restricted cluster. (B) Placenta cluster. (C) Digestive tract cluster. (D) Testis cluster.

Article Snippet: Secretome protein sequences without an InterPro or SCOP hit , 392.

Techniques: Gene Expression, Expressing, Hybridization, Labeling

Journal:

Article Title: The Mouse Secretome: Functional Classification of the Proteins Secreted Into the Extracellular Environment

doi: 10.1101/gr.983703

Figure Lengend Snippet: Properties of the 2033 Mouse Secretome Proteins

Article Snippet: Secretome protein sequences without an InterPro or SCOP hit , 392.

Techniques: Derivative Assay