Journal: Aging Cell

Article Title: Secretome Profiling of Young Multipotent Stem Cells Reveals Angiogenic and Immunomodulatory Mechanisms Supporting Aged Neuromuscular Health

doi: 10.1111/acel.70408

Figure Lengend Snippet: Systemic transplantation of young MDSPCs promotes skeletal muscle neovascularization, improves muscle structure, and increases mitochondrial content. (A) Representative immunohistochemical images of gastrocnemius (GS) muscles from young MDSPC (NA‐CI; n = 9) and PBS (NA‐PBS; n = 7) treated mice, labeled for CD31 (red) and dystrophin (green). (B) Quantification of vasculature density, presented as the number of vessels per muscle fiber. (C) Quantification of total vasculature area, presented as percentage of muscle area occupied by vessels. (D) Volcano plot depicting statistical significance and fold change of neovascularization pathway protein phosphorylation levels in GS muscles from NA‐CI ( n = 4) and NA‐PBS ( n = 4) mice. Proteins that have sites with significantly increased phosphorylation in NA‐CI muscles are highlighted in green. (E) Representative images of GS muscles stained with Sirius red (collagen, red) and Fast Green (muscle, green) at 2 months post‐intraperitoneal (IP) transplantation. (F) Quantification of collagen content as a percentage of total tissue area. (G) Representative images of dystrophin‐labeled GS muscles (green) from NA‐CI and NA‐PBS mice. (H) Violin plot of muscle fiber cross‐sectional area (CSA), with lines indicating median and interquartile ranges (25th and 75th percentiles). (I) Frequency distribution of muscle fiber CSA binned in 100 μm 2 intervals. (J) Citrate synthase activity measured in quadriceps muscles of NA‐CI ( n = 9) and NA‐PBS ( n = 7) mice. (K) Representative images of GS muscles labeled for type I (blue), type IIa (green), and type IIb (red) muscle fibers at 2 months post‐IP transplantation. (L) Stacked bar plot of fiber type composition in GS muscles of NA‐CI ( n = 7) and NA‐PBS ( n = 7) mice. Data (B, C, F, I, J, L) are presented as mean ± SEM. ** p ≤ 0.01, *** p ≤ 0.001, and § p ≤ 0.0001 using one‐tailed unpaired Student's t ‐test. (H) § p ≤ 0.0001 by two‐tailed Kolmogorov–Smirnov test. Scale bars are 100 μm (A, G, K) and 500 μm (E).

Article Snippet: Muscle fiber cross‐sectional area was assessed by immunohistochemically (IHC) labeling dystrophin (Abcam, ab15277, 1:300), type I muscle fibers (DSHB, BA‐F8, 1:50), type IIa muscle fibers (DSHB, SC‐71, 1:600), type IIb muscle fibers (DSHB, BF‐F3, 1:100), and type IIx muscle fibers (DSHB, 6H1, 1:50), following established protocols (Vella et al. ; Lavasani et al. , ; Bloemberg and Quadrilatero ).

Techniques: Transplantation Assay, Immunohistochemical staining, Muscles, Labeling, Phospho-proteomics, Staining, Activity Assay, One-tailed Test, Two Tailed Test

Journal: Aging Cell

Article Title: Secretome Profiling of Young Multipotent Stem Cells Reveals Angiogenic and Immunomodulatory Mechanisms Supporting Aged Neuromuscular Health

doi: 10.1111/acel.70408

Figure Lengend Snippet: Systemic transplantation of young MDSPCs promotes skeletal muscle neovascularization, improves muscle structure, and increases mitochondrial content. (A) Representative immunohistochemical images of gastrocnemius (GS) muscles from young MDSPC (NA‐CI; n = 9) and PBS (NA‐PBS; n = 7) treated mice, labeled for CD31 (red) and dystrophin (green). (B) Quantification of vasculature density, presented as the number of vessels per muscle fiber. (C) Quantification of total vasculature area, presented as percentage of muscle area occupied by vessels. (D) Volcano plot depicting statistical significance and fold change of neovascularization pathway protein phosphorylation levels in GS muscles from NA‐CI ( n = 4) and NA‐PBS ( n = 4) mice. Proteins that have sites with significantly increased phosphorylation in NA‐CI muscles are highlighted in green. (E) Representative images of GS muscles stained with Sirius red (collagen, red) and Fast Green (muscle, green) at 2 months post‐intraperitoneal (IP) transplantation. (F) Quantification of collagen content as a percentage of total tissue area. (G) Representative images of dystrophin‐labeled GS muscles (green) from NA‐CI and NA‐PBS mice. (H) Violin plot of muscle fiber cross‐sectional area (CSA), with lines indicating median and interquartile ranges (25th and 75th percentiles). (I) Frequency distribution of muscle fiber CSA binned in 100 μm 2 intervals. (J) Citrate synthase activity measured in quadriceps muscles of NA‐CI ( n = 9) and NA‐PBS ( n = 7) mice. (K) Representative images of GS muscles labeled for type I (blue), type IIa (green), and type IIb (red) muscle fibers at 2 months post‐IP transplantation. (L) Stacked bar plot of fiber type composition in GS muscles of NA‐CI ( n = 7) and NA‐PBS ( n = 7) mice. Data (B, C, F, I, J, L) are presented as mean ± SEM. ** p ≤ 0.01, *** p ≤ 0.001, and § p ≤ 0.0001 using one‐tailed unpaired Student's t ‐test. (H) § p ≤ 0.0001 by two‐tailed Kolmogorov–Smirnov test. Scale bars are 100 μm (A, G, K) and 500 μm (E).

Article Snippet: Muscle fiber cross‐sectional area was assessed by immunohistochemically (IHC) labeling dystrophin (Abcam, ab15277, 1:300), type I muscle fibers (DSHB, BA‐F8, 1:50), type IIa muscle fibers (DSHB, SC‐71, 1:600), type IIb muscle fibers (DSHB, BF‐F3, 1:100), and type IIx muscle fibers (DSHB, 6H1, 1:50), following established protocols (Vella et al. ; Lavasani et al. , ; Bloemberg and Quadrilatero ).

Techniques: Transplantation Assay, Immunohistochemical staining, Muscles, Labeling, Phospho-proteomics, Staining, Activity Assay, One-tailed Test, Two Tailed Test

Journal: Aging Cell

Article Title: Secretome Profiling of Young Multipotent Stem Cells Reveals Angiogenic and Immunomodulatory Mechanisms Supporting Aged Neuromuscular Health

doi: 10.1111/acel.70408

Figure Lengend Snippet: Systemic transplantation of young MDSPCs promotes skeletal muscle neovascularization, improves muscle structure, and increases mitochondrial content. (A) Representative immunohistochemical images of gastrocnemius (GS) muscles from young MDSPC (NA‐CI; n = 9) and PBS (NA‐PBS; n = 7) treated mice, labeled for CD31 (red) and dystrophin (green). (B) Quantification of vasculature density, presented as the number of vessels per muscle fiber. (C) Quantification of total vasculature area, presented as percentage of muscle area occupied by vessels. (D) Volcano plot depicting statistical significance and fold change of neovascularization pathway protein phosphorylation levels in GS muscles from NA‐CI ( n = 4) and NA‐PBS ( n = 4) mice. Proteins that have sites with significantly increased phosphorylation in NA‐CI muscles are highlighted in green. (E) Representative images of GS muscles stained with Sirius red (collagen, red) and Fast Green (muscle, green) at 2 months post‐intraperitoneal (IP) transplantation. (F) Quantification of collagen content as a percentage of total tissue area. (G) Representative images of dystrophin‐labeled GS muscles (green) from NA‐CI and NA‐PBS mice. (H) Violin plot of muscle fiber cross‐sectional area (CSA), with lines indicating median and interquartile ranges (25th and 75th percentiles). (I) Frequency distribution of muscle fiber CSA binned in 100 μm 2 intervals. (J) Citrate synthase activity measured in quadriceps muscles of NA‐CI ( n = 9) and NA‐PBS ( n = 7) mice. (K) Representative images of GS muscles labeled for type I (blue), type IIa (green), and type IIb (red) muscle fibers at 2 months post‐IP transplantation. (L) Stacked bar plot of fiber type composition in GS muscles of NA‐CI ( n = 7) and NA‐PBS ( n = 7) mice. Data (B, C, F, I, J, L) are presented as mean ± SEM. ** p ≤ 0.01, *** p ≤ 0.001, and § p ≤ 0.0001 using one‐tailed unpaired Student's t ‐test. (H) § p ≤ 0.0001 by two‐tailed Kolmogorov–Smirnov test. Scale bars are 100 μm (A, G, K) and 500 μm (E).

Article Snippet: Muscle fiber cross‐sectional area was assessed by immunohistochemically (IHC) labeling dystrophin (Abcam, ab15277, 1:300), type I muscle fibers (DSHB, BA‐F8, 1:50), type IIa muscle fibers (DSHB, SC‐71, 1:600), type IIb muscle fibers (DSHB, BF‐F3, 1:100), and type IIx muscle fibers (DSHB, 6H1, 1:50), following established protocols (Vella et al. ; Lavasani et al. , ; Bloemberg and Quadrilatero ).

Techniques: Transplantation Assay, Immunohistochemical staining, Muscles, Labeling, Phospho-proteomics, Staining, Activity Assay, One-tailed Test, Two Tailed Test