Journal: Nanomaterials

Article Title: Parenteral Nanoemulsions Loaded with Combined Immuno- and Chemo-Therapy for Melanoma Treatment

doi: 10.3390/nano12234233

Figure Lengend Snippet: MTT assay performed after 72 h of incubation with the formulations under study. B16 cells: upper left panel; D4M cells: upper right panel; M14 cells: lower left panel; A2058 cells: lower right panel. Abbreviations: IL: Intralipid ® 10%; SOR: sorafenib; TMZ: temozolomide; MIX: drug combination (temozolomide dodecyl ester, sorafenib, ICOS-Fc). Concentrations employed: SOR A = 16 μM; B = 10 μM; C = 8 μM; D = 4.5 μM; E = 0.8 μM. TMZ A = 48 μM; B = 32 μM; C = 24 μM; D = 16 μM; E = 2.4 μM. Statistical analysis: ^ p < 0.05 SOR vs. IL SOR; * p < 0.05 TMZ vs. IL TMZ; § p < 0.05 MIX vs. IL MIX.

Article Snippet: A2058 and M14 human melanoma cells and B16-F10 murine melanoma cells were purchased from the American Type Culture Collection (ATCC; Manassas, VA, USA).

Techniques: MTT Assay, Incubation

Journal: Nanomaterials

Article Title: Parenteral Nanoemulsions Loaded with Combined Immuno- and Chemo-Therapy for Melanoma Treatment

doi: 10.3390/nano12234233

Figure Lengend Snippet: Clonogenic assay with B16 (upper left panel), D4M (upper right panel), M14 (lower left panel), and A2058 (lower right panel) melanoma cells. Abbreviations: IL: Intralipid ® 10%; SOR: sorafenib; TMZ: temozolomide; MIX: drug combination (temozolomide dodecyl ester, sorafenib, ICOS-Fc). Cells were treated with the formulations under study for 3 h. Afterwards, the medium was changed, and cells were cultured in drug-free medium for an additional 7 days. Concentrations employed: SOR A = 16 μM; B = 10 μM; C = 8 μM; D = 4.5 μM; E = 0.8 μM. TMZ A = 48 μM; B = 32 μM; C = 24 μM; D = 16 μM; E = 2.4 μM. Statistical analysis: ^ p < 0.05 SOR vs. IL SOR; * p < 0.05 TMZ vs. IL TMZ; § p < 0.05 MIX vs. IL MIX.

Article Snippet: A2058 and M14 human melanoma cells and B16-F10 murine melanoma cells were purchased from the American Type Culture Collection (ATCC; Manassas, VA, USA).

Techniques: Clonogenic Assay, Cell Culture

Journal: Nanomaterials

Article Title: Parenteral Nanoemulsions Loaded with Combined Immuno- and Chemo-Therapy for Melanoma Treatment

doi: 10.3390/nano12234233

Figure Lengend Snippet: Hypothesized sorafenib (SOR) mechanism of action: ( a ) SOR release profile in culture medium, and ( b ) SOR internalization migration assay (B16, M14 melanoma cells): *** p < 0.005, * p < 0.05, # p < 0.01. ( c ) Hypothesized cellular pathways. Abbreviations: FCS: fetal calf serum; IL: Intralipid ® 10%; RAF: rapidly accelerated fibrosarcoma kinases; RTK: receptor tyrosine kinases.

Article Snippet: A2058 and M14 human melanoma cells and B16-F10 murine melanoma cells were purchased from the American Type Culture Collection (ATCC; Manassas, VA, USA).

Techniques: Migration

Journal: Stem Cells Translational Medicine

Article Title: Brachyury engineers cardiac repair competent stem cells

doi: 10.1002/sctm.20-0193

Figure Lengend Snippet: Brachyury (T) transfection provokes expression of cardiopoietic markers in human adipose‐derived mesenchymal stem cells (AMSCs). A, Immunofluorescent images demonstrate increased nuclear expressions of Nkx2.5 and Mef2c in AMSCs 72 hours post‐transfection with M 3 RNA encoding T (AMSC +T), compared to nontransfected AMSCs. Relative nuclear expressions of Nkx2.5 and Mef2c were compared based on fluorescence intensity normalized to nontransfected AMSCs. Numbers of biological replicates (n) were ≥17 for Nkx2.5 and ≥31 for Mef2c, per experiment condition. Scale bar = 20 μm. B, Increased levels of Nkx2.5 and Mef2c 72 hours post‐transfection were detected by real‐time quantitative polymerase chain reaction (n = 3). C, Western blot confirmed nuclear, not cytoplasmic, translocation of Nkx2.5 (n = 4) and Mef2c (n = 4) in AMSC +T at 72 hours post‐transfection. Protein expression was compared based on nuclear to cytoplasmic ratio of protein levels using (AMSC +T nuclear/AMSC +T cytoplasmic)/(AMSC nuclear/AMSC cytoplasmic) normalized to AMSC. Total protein stain was used as loading control. * P < .05; ** P < .01; *** P < .001 with Student's t test

Article Snippet: Using the M 3 RNA mRNA delivery platform, AMSCs were transfected with seven distinct mRNAs, namely myocyte enhancer factor 2C (Mef2c; TriLink, San Diego, California; M14‐AH02B), NK2 homeobox 5 (Nkx2.5; TriLink, M14‐AH01B), mesoderm posterior bHLH transcription factor 1 (Mesp1; TriLink, M14‐AK01A), Brachyury (TriLink, M14‐AN01A), octamer‐binding transcription factor 4 (Oct4; TriLink, T1‐APH01A), GATA binding protein 4 (Gata4; TriLink, M14‐AH04A), and T‐box transcription factor 5 (Tbx5; TriLink, M14‐AH03A), individually or in various combinations.

Techniques: Transfection, Expressing, Derivative Assay, Fluorescence, Real-time Polymerase Chain Reaction, Western Blot, Translocation Assay, Staining

Journal: Stem Cells Translational Medicine

Article Title: Brachyury engineers cardiac repair competent stem cells

doi: 10.1002/sctm.20-0193

Figure Lengend Snippet: Brachyury (T) transforms the adipose‐derived mesenchymal stem cell transcriptome. A, Heat map was generated using RNA sequencing data from cell lysates at five time points; 0‐ (pre), 24‐, 48‐, 72‐, and 96‐hours after T‐transfection. Each time point consisted of biological triplicates (n = 3). Quantile‐adjusted conditional maximum likelihood method (qCML) was used for the initial bioinformatics analysis. B, Gene Ontology analysis of differentially expressed genes (upregulated at 72 hours vs 0 hour, based on P < .05 and log2‐fold change >1 cutoff) identified biological processes with enriched representation. T‐induced transcriptome prioritization included antioxidant/response to stimuli (green); angiogenesis/circulatory development (red); immunomodulatory/anti‐inflammatory response (yellow); cell, tissue, and organ development/differentiation (blue). These biological functions are all considered to be of potential benefit in cardiac repair

Article Snippet: Using the M 3 RNA mRNA delivery platform, AMSCs were transfected with seven distinct mRNAs, namely myocyte enhancer factor 2C (Mef2c; TriLink, San Diego, California; M14‐AH02B), NK2 homeobox 5 (Nkx2.5; TriLink, M14‐AH01B), mesoderm posterior bHLH transcription factor 1 (Mesp1; TriLink, M14‐AK01A), Brachyury (TriLink, M14‐AN01A), octamer‐binding transcription factor 4 (Oct4; TriLink, T1‐APH01A), GATA binding protein 4 (Gata4; TriLink, M14‐AH04A), and T‐box transcription factor 5 (Tbx5; TriLink, M14‐AH03A), individually or in various combinations.

Techniques: Derivative Assay, Generated, RNA Sequencing Assay, Transfection

Journal: Stem Cells Translational Medicine

Article Title: Brachyury engineers cardiac repair competent stem cells

doi: 10.1002/sctm.20-0193

Figure Lengend Snippet: Engineered cardiopoietic stem cells (eCP) demonstrate elevated antioxidant capacity. A, Oxidative stress was induced in AC16 human cardiomyocytes using the free‐radical generator LY83583. Four treatment groups included; culture medium alone (Media), plus OptiMEM (+OptiMEM), plus adipose‐derived mesenchymal stem cell‐derived conditioned media (+AMSC CM), or plus Brachyury transfected engineered cardiopoietic stem cell‐derived conditioned media (+eCP CM). In each treatment group, 20 μL of conditioned media was added to 200 μL of regular media per well. Caspase 3/7 Green and NucLight Red identified apoptotic cells and cell nuclei, respectively (top panels). Significant decrease in apoptotic cardiomyocytes (bottom panels) indicated a protective effect of eCP conditioned media. Numbers of biological replicates (n) were nine per group at each time point. Scale bar = 100 μm. Antioxidant capacity was consistently detected as measured by: expression of antioxidant proteins SOD2 and SOD3 in total cell lysate (Western blot, n = 3; B); superoxide dismutase (SOD) activity in conditioned media (n = 3; C); and total antioxidant capacity in conditioned media (n = 3; D). OptiMEM served as control. * P < .05; ** P < .01; *** P < .001 with one‐way ANOVA followed by a post hoc Bonferroni test (A, C, D) or Student's t test (B). ANOVA, analysis of variance

Article Snippet: Using the M 3 RNA mRNA delivery platform, AMSCs were transfected with seven distinct mRNAs, namely myocyte enhancer factor 2C (Mef2c; TriLink, San Diego, California; M14‐AH02B), NK2 homeobox 5 (Nkx2.5; TriLink, M14‐AH01B), mesoderm posterior bHLH transcription factor 1 (Mesp1; TriLink, M14‐AK01A), Brachyury (TriLink, M14‐AN01A), octamer‐binding transcription factor 4 (Oct4; TriLink, T1‐APH01A), GATA binding protein 4 (Gata4; TriLink, M14‐AH04A), and T‐box transcription factor 5 (Tbx5; TriLink, M14‐AH03A), individually or in various combinations.

Techniques: Derivative Assay, Transfection, Expressing, Western Blot, Activity Assay