Journal: Stem Cell Research & Therapy

Article Title: Human bone marrow-derived, pooled, allogeneic mesenchymal stromal cells manufactured from multiple donors at different times show comparable biological functions in vitro, and in vivo to repair limb ischemia

doi: 10.1186/s13287-021-02330-9

Figure Lengend Snippet: Gene expression profiling. a Unsupervised hierarchical clustering/condition tree of samples, HFF-1, three batches of Stempeucel®-1 and two batches of Stempeucel®-1A together based on the similarity of their expression profiles. It is evident from the condition tree based on the distance that out of 24,498 genes, an average of ≥ 77% of the global profiles of all the Stempeucel® products is similar compared to the HFF cells indicating a passage-independent similarity in global gene expression profiles. ~ 20% of the genes (4924 out of 24,498 genes) analyzed were differentially expressed between Stempeucel®-1 and Stempeucel®-1A . In total, ~ 3% of the genes (722 out of 24,498 genes analyzed) were significantly differentially expressed with a fold change of ≥ 2. b Analysis of genes expressed by Stempeucel®-1 ( X -axis) and Stempeucel®-1A (Y-axis) with a significantly high correlation of R 2 = 0.9827 in a scatter plot. c Comparative evaluation of twenty-three angiogenic genes in HFF-1 cell line, three batches of Stempeucel®-1, and two batches of Stempeucel®-1A. d Real-time PCR analysis of five selected angiogenic genes co-expressed across the Stempeucel® batches compared to HFF-1; VEGF—* P = 0.03, AGPT1—* P = 0.04, TGF b1—* P = 0.01, HGF— P = 0.03, and CXCL8—* P = 0.02. e Comparative evaluation of thirty-four anti-inflammation genes in HFF-1, three batches of Stempeucel®-1 and two batches of Stempeucel®-1A. f Real-time PCR analysis of five selected anti-inflammatory genes co-expressed across Stempeucel® batches compared to HFF-1; IDO—** P = 0.001, IL-10—** P = 0.004, VCAM1 ** P = 0.002, PDL1—** P = 0.009, and HLA-G—** P = 0.002

Article Snippet: Stempeucel®-1 is formulated and cryopreserved in 85% Plasmalyte A (Baxter, Illinois, USA), 10% dimethyl sulfoxide (DMSO) (Sigma-Aldrich, Missouri, USA), and 5% human serum albumin (HSA) (Sigma-Aldrich, Missouri, USA), and Stempeucel®-1A is formulated in CS5 (CryoStor 5, Biolife Solutions, Washington, USA).

Techniques: Expressing, Real-time Polymerase Chain Reaction

Journal: Stem Cell Research & Therapy

Article Title: Human bone marrow-derived, pooled, allogeneic mesenchymal stromal cells manufactured from multiple donors at different times show comparable biological functions in vitro, and in vivo to repair limb ischemia

doi: 10.1186/s13287-021-02330-9

Figure Lengend Snippet: In vitro immunosuppression of alloreactive MLRs by Stempeucel®-1 and Stempeucel®-1A. a Suppression of MLR by mitomycin C-arrested Stempeucel®-1 ( n = 15) and Stempeucel®-1A ( n = 4) at three MLR: MSC ratios of 1:1, 1:2.5, and 1:10; MLR proliferation and suppression was measured by BrdU incorporation measurement as described in the “ ” section. b Percentage of cells positive for HLA-DR in Stempeucel®-1 ( n = 3) and Stempeucel®-1A ( n = 3) cells at the basal level and after priming with TNFα and IFN-γ showed significant difference (*** P = 0.0007). c Immunosuppression activity was retained in both Stempeucel®-1 and Stempeucel®-1A upon inflammatory priming at all three ratios of IFN-γ and TNFα primed, high HLA-DR expressing Stempeucel®-1 and Stempeucel®-1A cells did not reduce their immunosuppression capacity and a trend towards higher immunosuppression by primed cells was seen compared to unprimed controls. Graphs represent mean ± SD values; ns – not significant

Article Snippet: Stempeucel®-1 is formulated and cryopreserved in 85% Plasmalyte A (Baxter, Illinois, USA), 10% dimethyl sulfoxide (DMSO) (Sigma-Aldrich, Missouri, USA), and 5% human serum albumin (HSA) (Sigma-Aldrich, Missouri, USA), and Stempeucel®-1A is formulated in CS5 (CryoStor 5, Biolife Solutions, Washington, USA).

Techniques: In Vitro, BrdU Incorporation Assay, Activity Assay, Expressing

Journal: Stem Cell Research & Therapy

Article Title: Human bone marrow-derived, pooled, allogeneic mesenchymal stromal cells manufactured from multiple donors at different times show comparable biological functions in vitro, and in vivo to repair limb ischemia

doi: 10.1186/s13287-021-02330-9

Figure Lengend Snippet: Quantification of angiogenic growth factors. a VEGF (A), b SDF-1α, c IL-8, and d TGFβ1 levels in the CM collected at the 72 h time point from Stempeucel®-1 and Stempeucel®-1A batches. Significant variation in the levels was observed between the two Stempeucel® versions, VEGF—** P = 0.001; SDF-1α—* P = 0.01; IL-8—*** P = 0.0006; TGFβ1—** P = 0.003. e The CMs derived from both Stempeucel®-1 and Stempeucel®-1A promoted HUVEC migration equally. Addition of anti-VEGF mAb in the CMs significantly inhibited HUVEC migration (Stempeucel®-1 = 61.6%; Stempeucel®-1A = 61.6%). f Equivalent HUVEC proliferation was observed with both Stempeucel®-derived CM. Significant inhibition in HUVEC proliferation was observed upon neutralizing VEGF in the CMs (Stempeucel®-1 = 97.9, Stempeucel®-1A = 100%). g No significant difference in the HUVEC tube formation efficiency was observed between the CMs derived from Stempeucel®-1 and Stempeucel®-1A and similar inhibition (Stempeucel®-1 = 69.6%, Stempeucel®-1A = 65.1%) was observed between the two CMs in the presence of anti-VEGF mAb; a, b Similar HUVEC tube forming efficiency was observed with the CMs from both Stempeucel®-1 and Stempeucel®-1A. EGM—endothelial growth medium control; KO + 10%FBS – DMEM-KO basal medium plus 10% FBS control; SFM: serum-free DMEM-KO medium. Graphs represent mean ± SEM values

Article Snippet: Stempeucel®-1 is formulated and cryopreserved in 85% Plasmalyte A (Baxter, Illinois, USA), 10% dimethyl sulfoxide (DMSO) (Sigma-Aldrich, Missouri, USA), and 5% human serum albumin (HSA) (Sigma-Aldrich, Missouri, USA), and Stempeucel®-1A is formulated in CS5 (CryoStor 5, Biolife Solutions, Washington, USA).

Techniques: Derivative Assay, Migration, Inhibition

Journal: Stem Cell Research & Therapy

Article Title: Human bone marrow-derived, pooled, allogeneic mesenchymal stromal cells manufactured from multiple donors at different times show comparable biological functions in vitro, and in vivo to repair limb ischemia

doi: 10.1186/s13287-021-02330-9

Figure Lengend Snippet: A similar magnitude of reduction in muscle degeneration and improvement in angiogenesis in the ischemic limbs of mice following Stempeucel®-1 and Stempeucel®-1A treatment: a Treatment with Stempeucel®-1 and Stempeucel®-1A significantly protected muscle fiber loss of adductor and gastrocnemius muscles compared to vehicle control. b Protection from muscle weight loss was observed upon treatment with Stempeucel®-1 and Stempeucel®-1A compared to the vehicle-treated group. c–f Histological analysis of muscle section of the sham control, vehicle control, and Stempeucel-treated groups. Red arrow indicates vacuolar degeneration, green arrow—mononuclear cells infiltration. Yellow arrow—muscle necrosis and black arrow—muscle degeneration. Intramuscular administrations of Stempeucel®-1 and Stempeucel®-1A survive in ischemic limb tissue, secrete paracrine factors, and recruit or proliferate CD31 positive endothelial cells. Representative images of IHC staining for g–j mCD31; k–n HNA; o–r hVEGF; positive cells in limb muscles of sham control, vehicle control, and Stempeucel-treated animals at day 28. Positive areas are marked with black arrows. Photographs were taken at × 40 magnification, and the scale bar shows the 50 μm

Article Snippet: Stempeucel®-1 is formulated and cryopreserved in 85% Plasmalyte A (Baxter, Illinois, USA), 10% dimethyl sulfoxide (DMSO) (Sigma-Aldrich, Missouri, USA), and 5% human serum albumin (HSA) (Sigma-Aldrich, Missouri, USA), and Stempeucel®-1A is formulated in CS5 (CryoStor 5, Biolife Solutions, Washington, USA).

Techniques: Immunohistochemistry

Journal: Stem Cell Research & Therapy

Article Title: Human bone marrow-derived, pooled, allogeneic mesenchymal stromal cells manufactured from multiple donors at different times show comparable biological functions in vitro, and in vivo to repair limb ischemia

doi: 10.1186/s13287-021-02330-9

Figure Lengend Snippet: Stempeucel®-1 and Stempeucel®-1A treatment improved blood perfusion and capillary density in ischemic limbs. a, b Laser Doppler blood flow flux was measured in sham, LI + vehicle, and LI + two Stempeucel®-treated groups at 28 days after LI. ## P < 0.01, sham control vs LI vehicle; ** P < 0.01, LI vehicle vs LI + Stempeucel®-1; * P < 0.05, LI vehicle vs LI + Stempeucel®-1A. c Capillary density at day 28 increased significantly in Stempeucel®-1- (** P < 0.001) and Stempeucel®-1A (** P < 0.01)-treated animals compared to vehicle-treated animals. Values are expressed as mean ± SEM. Scale bar = 50 μm, × 40 magnification

Article Snippet: Stempeucel®-1 is formulated and cryopreserved in 85% Plasmalyte A (Baxter, Illinois, USA), 10% dimethyl sulfoxide (DMSO) (Sigma-Aldrich, Missouri, USA), and 5% human serum albumin (HSA) (Sigma-Aldrich, Missouri, USA), and Stempeucel®-1A is formulated in CS5 (CryoStor 5, Biolife Solutions, Washington, USA).

Techniques:

Journal: Stem Cell Research & Therapy

Article Title: Human bone marrow-derived, pooled, allogeneic mesenchymal stromal cells manufactured from multiple donors at different times show comparable biological functions in vitro, and in vivo to repair limb ischemia

doi: 10.1186/s13287-021-02330-9

Figure Lengend Snippet: Biodistribution kinetics of Stempeucel®-1 in sham and ischemia-induced BALB/c nude mice. a Heat map of DiI-labeled areas in the limb muscle of sham control and ischemic animals (one representative animal shown) at various time points after Stempeucel®-1 injection. RGB scale—1 × 10 7 –10 × 10 7 photons/s/mm 2 . b Graphical representation of the average net intensity of DiI-labeled cells in the limb muscles of sham and ischemic animal groups on day 1, day 3, day 6, day 11, day 14, day 21, and day 28 (end of the experiment). Significantly higher intensity of DiI signal was observed in the ischemia-induced animal group on day 6 and 11 (* P = 0.01) compared to that of the sham group

Article Snippet: Stempeucel®-1 is formulated and cryopreserved in 85% Plasmalyte A (Baxter, Illinois, USA), 10% dimethyl sulfoxide (DMSO) (Sigma-Aldrich, Missouri, USA), and 5% human serum albumin (HSA) (Sigma-Aldrich, Missouri, USA), and Stempeucel®-1A is formulated in CS5 (CryoStor 5, Biolife Solutions, Washington, USA).

Techniques: Labeling, Injection