Journal: Scientific Reports

Article Title: Highly variable biological effects of statins on cancer, non-cancer, and stem cells in vitro

doi: 10.1038/s41598-024-62615-w

Figure Lengend Snippet: The effect of statins on viability and growth of ( a ) stem ADMSC and non-cancerous HEK 293 cells and ( b ) cancer MiaPaCa-2 cells. ( a ) ADMSC—human adipose-derived mesenchymal stem cells, HEK 293—human embryonic kidney cells, exposure to statins—24 h, concentrations 0—100 µM, control—methanol, ( b ) previously published data , MiaPaCa-2—pancreatic cancer cells, exposure to statins—24 h, concentrations 0—40 µM, control—methanol.

Article Snippet: Human adipose-derived mesenchymal stem cells ADMSC (ATCC, Manassas, VA, PSC-500-011, LOT 70017032, positive specific staining for CD29, CD44, CD73, CD90, CD105, and CD166 and negative for CD14, CD31, CD34, and CD45) were cultured in mesenchymal stem cell basal medium (ATCC, Manassas, VA) supplemented with low serum mesenchymal stem cell growth kit for adipose- and umbilical-derived MSCs (ATCC, Manassas, VA).

Techniques: Derivative Assay, Control

Journal: Scientific Reports

Article Title: Highly variable biological effects of statins on cancer, non-cancer, and stem cells in vitro

doi: 10.1038/s41598-024-62615-w

Figure Lengend Snippet: Comparison of the effect of statins on the growth and viability of pancreatic cancer MiaPaCa-2 cells, non-cancerous HEK 293 cells, and ADMSC stem cells. Concentration of statins—20 µM, Time—exposure to statins—24, 48, and 72 h, control—methanol.

Article Snippet: Human adipose-derived mesenchymal stem cells ADMSC (ATCC, Manassas, VA, PSC-500-011, LOT 70017032, positive specific staining for CD29, CD44, CD73, CD90, CD105, and CD166 and negative for CD14, CD31, CD34, and CD45) were cultured in mesenchymal stem cell basal medium (ATCC, Manassas, VA) supplemented with low serum mesenchymal stem cell growth kit for adipose- and umbilical-derived MSCs (ATCC, Manassas, VA).

Techniques: Comparison, Concentration Assay, Control

Journal: Scientific Reports

Article Title: Highly variable biological effects of statins on cancer, non-cancer, and stem cells in vitro

doi: 10.1038/s41598-024-62615-w

Figure Lengend Snippet: Effect of statins on size and compactness of spheroids. ( a ) ADMSC stem cells, ( b ) pancreatic cancer MiaPaCa-2 cells, concentration of statins—20 µM, Ctr—methanol treated spheroids, P—pravastatin, R—rosuvastatin, L—lovastatin, F—fluvastatin, A—atorvastatin, Pi—pitavastatin, C—cerivastatin, S—simvastatin. Statins were added once, after spheroid formation, 10 weeks ( a ) or 3.5 weeks ( b ) after inoculation. Experiment was carried out in biological dodecaplicates.

Article Snippet: Human adipose-derived mesenchymal stem cells ADMSC (ATCC, Manassas, VA, PSC-500-011, LOT 70017032, positive specific staining for CD29, CD44, CD73, CD90, CD105, and CD166 and negative for CD14, CD31, CD34, and CD45) were cultured in mesenchymal stem cell basal medium (ATCC, Manassas, VA) supplemented with low serum mesenchymal stem cell growth kit for adipose- and umbilical-derived MSCs (ATCC, Manassas, VA).

Techniques: Concentration Assay

Journal: Scientific Reports

Article Title: Highly variable biological effects of statins on cancer, non-cancer, and stem cells in vitro

doi: 10.1038/s41598-024-62615-w

Figure Lengend Snippet: Effect of statins on the spheroid formation. ( a ) ADMSC stem cells, ( b ) pancreatic cancer MiaPaCa-2 cells, concentration of statins—20 µM, Ctr methanol treated spheroids, P —pravastatin, R —rosuvastatin, L —lovastatin, F —fluvastatin, A —atorvastatin, Pi —pitavastatin, C —cerivastatin, S —simvastatin. Statins were added once, 24 h after cell inoculation. Experiment was carried out in biological dodecaplicates.

Article Snippet: Human adipose-derived mesenchymal stem cells ADMSC (ATCC, Manassas, VA, PSC-500-011, LOT 70017032, positive specific staining for CD29, CD44, CD73, CD90, CD105, and CD166 and negative for CD14, CD31, CD34, and CD45) were cultured in mesenchymal stem cell basal medium (ATCC, Manassas, VA) supplemented with low serum mesenchymal stem cell growth kit for adipose- and umbilical-derived MSCs (ATCC, Manassas, VA).

Techniques: Concentration Assay

Journal: Scientific Reports

Article Title: Highly variable biological effects of statins on cancer, non-cancer, and stem cells in vitro

doi: 10.1038/s41598-024-62615-w

Figure Lengend Snippet: Comparison of expression changes between statin treated and control MiaPaCa-2 and ADMSC cells. Displayed are only the genes that are differentially expressed upon at least one statin treatment in at least one cell type, requiring |log 2 FC|> 1 and FDR < 0.05. Statins were administered at a concentration of 12 µM for 24 h. ( FC fold change, FDR false discovery rate, horizontal and vertical axes—changes in ADMSC and MiaPaCa-2 cells, respectively, upon respective treatment). The red dashed lines indicate two-fold change increase or decrease in the gene expression. The genes with at least two-fold up-regulation (resp. down-regulation) in ADMSC stem cells are displayed to the right (resp. left) of the dashed lines. Similarly, genes with at least two-fold up-regulation (resp. down-regulation) in cancer cells are displayed above (resp. below) of the dashed lines. For details about differentially regulated transcripts see the ArrayExpress database, accessions E-MTAB-3979, E-MTAB-11579 .

Article Snippet: Human adipose-derived mesenchymal stem cells ADMSC (ATCC, Manassas, VA, PSC-500-011, LOT 70017032, positive specific staining for CD29, CD44, CD73, CD90, CD105, and CD166 and negative for CD14, CD31, CD34, and CD45) were cultured in mesenchymal stem cell basal medium (ATCC, Manassas, VA) supplemented with low serum mesenchymal stem cell growth kit for adipose- and umbilical-derived MSCs (ATCC, Manassas, VA).

Techniques: Comparison, Expressing, Control, Concentration Assay, Gene Expression

Journal: Scientific Reports

Article Title: Highly variable biological effects of statins on cancer, non-cancer, and stem cells in vitro

doi: 10.1038/s41598-024-62615-w

Figure Lengend Snippet: Cellular pathways most significantly affected by statins in cancer and stem cells. The gene set enrichment analysis (GSEA) revealed the KEGG pathways most affected by statin treatment in ADMSC and MiaPaCa-2 cells. Displayed is the union of the top five most enriched pathways among the comparisons. (Statin concentration—12 µM, treatment time—24 h, p-value—GSEA p-value, gene ratio—fraction of KEGG pathway genes among differentially expressed genes). For details about differentially regulated transcripts, see the ArrayExpress database, accessions E-MTAB-3979, E-MTAB-11579.

Article Snippet: Human adipose-derived mesenchymal stem cells ADMSC (ATCC, Manassas, VA, PSC-500-011, LOT 70017032, positive specific staining for CD29, CD44, CD73, CD90, CD105, and CD166 and negative for CD14, CD31, CD34, and CD45) were cultured in mesenchymal stem cell basal medium (ATCC, Manassas, VA) supplemented with low serum mesenchymal stem cell growth kit for adipose- and umbilical-derived MSCs (ATCC, Manassas, VA).

Techniques: Concentration Assay

Journal: mAbs

Article Title: Assessment of the role of afucosylated glycoforms on the in vitro antibody-dependent phagocytosis activity of an antibody to Aβ aggregates

doi: 10.1080/19420862.2020.1803645

Figure Lengend Snippet: Antibody-mediated Aβ uptake in human cell systems. (a-c) Mean number of Fc Receptors determined using the QuantiBRITE PE kit on two different human macrophage preparations derived from human peripheral CD14+ monocytes (A, n = 1–2) and on the human THP-1 cell line (B, n = 2). Mean ± S.E.M. are provided. (c) Kifunensine high mannose sample (HM-Afuc) has similar dose-dependent Aβ phagocytosis activity as the standard mediated by primary human macrophages (two-way ANOVA for standard vs sample, p >.05; n = 3). Results are presented as the relative ADCP fluorescence unit (RFU) as measured using the trypan blue exclusion method versus antibody concentration. Mean ± S.E.M. and non-linear regression curve fit analysis are shown. (d) HM-Afuc has similar dose-dependent Aβ phagocytosis activity as the standard mediated by the THP-1 cell line (two-way ANOVA for standard vs sample, p >.05; n = 4). Samples were run in duplicate and Aβ-GFP phagocytosis was quantitated by flow cytometry. The mean ± S.E.M. of the percent positive GFP cells of 1000 cells sampled per sample and non-linear regression curve-fit analysis are shown.

Article Snippet: Human peripheral blood CD14+ monocytes (70035, Stem Cell Technologies) were differentiated to macrophages with recombinant M-CSF (216-MC, R&D Systems) using the following media (Advanced RPMI1640 medium, 5% FBS, Penicillin-Streptomycin 1x, L-Glutamine 2 mM, M-CSF 100 ng/ml).

Techniques: Derivative Assay, Activity Assay, Fluorescence, Concentration Assay, Flow Cytometry

Journal:

Article Title: Reassessing the role of HLA-DRB3 T cell responses: Evidence for significant expression and complementary antigen presentation

doi: 10.1002/eji.200939225

Figure Lengend Snippet: Level of DRB1-52 and DRB3 in CD19+ (A) and CD14+ (B) cells (see Supporting Information Table 1 for donor information). Data are expressed as ratio of DRB1 (grey) or DRB3 (black) to GAPDH × 105 copy numbers, and referred to as RQ. Data show mean + SD (n=3). I: donor with DRB1*0301/*1301 - DRB3*0101/*0101 typing; II: donors with DRB1*0301 and DRB3*0101 typing; III: donors with DRB1*1301 and DRB3*0101 typing; IV: donors with typing DRB1*1101 and DRB3*0202 typing. The paired comparison (two tailored Wilcoxon signed test) of DRB1 and DRB3 mRNA in both cell populations indicated that DRB1-52 is expressed at significantly higher levels (p=0.0078) than DRB3.

Article Snippet: 4.2 CD19+ and CD14+ cell separation, RNA extraction and Reverse transcription From PBMC, monocytes and B cells were purified by positive selection using Human CD14 Selection Cocktail (Stem Cell Technologies) and CD19 MicroBeads (Miltenyi, Auburn, CA), respectively.

Techniques: Comparison

Journal:

Article Title: Reassessing the role of HLA-DRB3 T cell responses: Evidence for significant expression and complementary antigen presentation

doi: 10.1002/eji.200939225

Figure Lengend Snippet: The observed MFI for DRB3 antibody staining in seven donors is plotted for CD19+ cells (left), CD14+ cells (middle) with mean values indicated by horizontal lines. The CD19+/CD14+ ratio (right) is also plotted. Symbols in all the representations stand for donors typing, open circles for DRB1*13-DRB3*0101, filled triangles for DRB1*11-DRB3*0202, filled squares for DRB1*03-DRB3*0101 and filled circles for DRB1*03/13-DRB3*0101/0101. The level of DRB3 MFI was higher on B cells than on monocytes, p=0.0148 (two tailored Wilcoxon signed test).

Article Snippet: 4.2 CD19+ and CD14+ cell separation, RNA extraction and Reverse transcription From PBMC, monocytes and B cells were purified by positive selection using Human CD14 Selection Cocktail (Stem Cell Technologies) and CD19 MicroBeads (Miltenyi, Auburn, CA), respectively.

Techniques: Staining

Journal: Cell reports

Article Title: Macrophage memories of early-life injury drive neonatal nociceptive priming

doi: 10.1016/j.celrep.2024.114129

Figure Lengend Snippet: (A) Example protein arrays of BMDM lysates with knockdown of p75NTR and stimulated with the indicated factors. (B) Heatmap indicating protein expression detected across each stimulus; n = 4/group. (C) Examples of significantly regulated proteins from the arrays (* p < 0.05 vs. siCON+Veh, ^ p < 0.05 vs. sip75+LPS, # p = 0.05 vs. sip75+Veh, ## p > 0.05 vs. sip75+Veh; one-way ANOVA, Tukey’s or ANOVA on ranks, Dunn’s; n = 4/group). (D–F) Representative images of iPSC-derived macrophages, sensory neurons, and Rhod-2 responses to ATP. Arrows, increased responses; arrowheads, new responses. Scale bar, 25 μm. (G) Example calcium transients from iPSC-derived sensory neurons in response to ATP, capsaicin, or KCl. Arrows indicate when the ATP or capsaicin was added, and the brackets indicate the part of the trace that was analyzed to calculate responses. scale bar, 20 μM. (H) Exposure of iPSC-derived sensory neurons to medium from macrophages with loss of p75NTR treated with NGF caused decreased responsiveness of neurons to ATP (F = 4.257) (* p < 0.04 vs. all other conditions, one-way ANOVA with Tukey’s post hoc, n = 15 cells per group). (I) No changes in capsaicin responses (F = 2.702) were observed (one way ANOVA). Mean ± SEM or percent change from controls with variance.

Article Snippet: Human induced pluripotent stem cells (iPSC) derived CD14 + monocytes (ATCC, Manassas, VA, Cat No. DYS0100) were plated in macrophage differentiation medium (RPMI-1640 Medium/10% Fetal Bovine Serum (FBS), supplemented with 100 ng/mL human M-CSF, and 1% Pen/Strep) in 24 well plate at a density of 10 ^ 5 cells/ml according to manufacturer’s directions.

Techniques: Knockdown, Expressing, Derivative Assay

Journal: Cell reports

Article Title: Macrophage memories of early-life injury drive neonatal nociceptive priming

doi: 10.1016/j.celrep.2024.114129

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Human induced pluripotent stem cells (iPSC) derived CD14 + monocytes (ATCC, Manassas, VA, Cat No. DYS0100) were plated in macrophage differentiation medium (RPMI-1640 Medium/10% Fetal Bovine Serum (FBS), supplemented with 100 ng/mL human M-CSF, and 1% Pen/Strep) in 24 well plate at a density of 10 ^ 5 cells/ml according to manufacturer’s directions.

Techniques: Recombinant