mouse brain endothelial cells bend3  (ATCC)

Journal: ACS Applied Bio Materials

Article Title: Real-Time Monitoring of the Cytotoxic Effect of Oxygen-Sensitive Fluorescent Poly(styrene-maleic anhydride) Nanoparticles Using Electrical-Substrate Impedance Sensing

doi: 10.1021/acsabm.5c01443

Figure Lengend Snippet: Effects of SMA NPs on viable cell density and toxicity of NIH 3T3 fibroblasts and bEnd3 brain endothelial cells assessed by alamarBlue assay: (a–c) NIH 3T3 fibroblasts; (d–f) bEnd3 cells. (a) Fluorescence intensity of reduced resazurin by NIH 3T3 fibroblasts on day 5 of culture corresponding to 3 days of incubation with SMA NPs. (b) Relative cell density of NIH 3T3 fibroblasts on day 5. (c) Cytotoxicity of SMA NPs on NIH 3T3 fibroblasts. (d) Fluorescence intensity of reduced resazurin by bEnd3 cells on day 5 of culture corresponding to 3 days of incubation with SMA NPs. (e) Relative cell density of bEnd3 cells on day 5. (f) Cytotoxicity of SMA NPs on bEnd3 cells. In all panels, the fluorescence intensity is compared to the negative control (brown bars). **** p < 0.0001. n = 4.

Article Snippet: bEnd3 murine brain endothelial cells, obtained from the American Type Culture Collection (ATCC, CRL-2299, VA), were cultured in Gibco DMEM (low glucose) supplemented with 10% fetal bovine serum (Sigma-Aldrich), 0.1% gentamicin (Thermo Fisher Scientific).

Techniques: Alamar Blue Assay, Fluorescence, Incubation, Negative Control

Journal: ACS Applied Bio Materials

Article Title: Real-Time Monitoring of the Cytotoxic Effect of Oxygen-Sensitive Fluorescent Poly(styrene-maleic anhydride) Nanoparticles Using Electrical-Substrate Impedance Sensing

doi: 10.1021/acsabm.5c01443

Figure Lengend Snippet: ECIS measurements of cellular response to the introduction of fluorescent SMA NPs. (a) Normalized five-day real-time ECIS measurement of NIH 3T3 fibroblasts, showing real-time data of cell attachment, proliferation, and response to fluorescent SMA NPs at varying concentrations. Capacitance was recorded at 64 kHz. The dotted blue line indicates the time when the fluorescent SMA NPs were added to each well. (b) Capacitance values from NIH 3T3 cells on day five at 110 h in culture. (c) Cytotoxicity percentage of NIH 3T3 fibroblasts in response to contact with SMA NPs. (d) Normalized five-day real-time ECIS measurement of bEnd3brain epithelial cells, showing real-time data of attachment, proliferation, and response to fluorescent SMA NPs. Capacitance was recorded at 64 kHz. The dotted blue line indicates the time when the fluorescent SMA NPs were added to each well. (e) Capacitance values of bEnd3 cells on day five at 110 h in culture. (f) Cytotoxicity percentage of brain epithelial bEnd3 cells in response to contact with SMA NPs. In panels b, c, e, and f, the capacitances and cytotoxicities are compared to the negative control (brown bars). **** p < 0.0001. n = 4.

Article Snippet: bEnd3 murine brain endothelial cells, obtained from the American Type Culture Collection (ATCC, CRL-2299, VA), were cultured in Gibco DMEM (low glucose) supplemented with 10% fetal bovine serum (Sigma-Aldrich), 0.1% gentamicin (Thermo Fisher Scientific).

Techniques: Cell Attachment Assay, Negative Control

Journal: ACS Applied Bio Materials

Article Title: Real-Time Monitoring of the Cytotoxic Effect of Oxygen-Sensitive Fluorescent Poly(styrene-maleic anhydride) Nanoparticles Using Electrical-Substrate Impedance Sensing

doi: 10.1021/acsabm.5c01443

Figure Lengend Snippet: Barrier condition analysis of bENd3 brain epithelial cells after addition of SMA NPs at different concentrations. (a) Real-time ECIS resistance measurements at 4 kHz. (b) Comparison of resistance values between different experimental conditions, before adding SMA NPs on day 4, after SMA NPs addition on day 8, and after media replacement on day 10. (c) Real-time ECIS capacitance measurements at 64 kHz. (d) Comparison of normalized capacitance between different experimental conditions, before adding SMA NPs, after SMA NPs addition, and after media replacement. In panels b and d, the capacitance is compared to the negative control (brown bars). ** p < 0.0001. n = 5.

Article Snippet: bEnd3 murine brain endothelial cells, obtained from the American Type Culture Collection (ATCC, CRL-2299, VA), were cultured in Gibco DMEM (low glucose) supplemented with 10% fetal bovine serum (Sigma-Aldrich), 0.1% gentamicin (Thermo Fisher Scientific).

Techniques: Comparison, Negative Control

Journal: ACS Applied Bio Materials

Article Title: Real-Time Monitoring of the Cytotoxic Effect of Oxygen-Sensitive Fluorescent Poly(styrene-maleic anhydride) Nanoparticles Using Electrical-Substrate Impedance Sensing

doi: 10.1021/acsabm.5c01443

Figure Lengend Snippet: Fluorescence-phase contrast micrographs of randomly selected samples. Micrographs show NIH 3T3 fibroblasts (top panel) and bEnd3 brain endothelial cells (bottom panel) containing fluorescent SMA NPs (green) on day 7 at multiple concentrations. Scale bar = 275 μm.

Article Snippet: bEnd3 murine brain endothelial cells, obtained from the American Type Culture Collection (ATCC, CRL-2299, VA), were cultured in Gibco DMEM (low glucose) supplemented with 10% fetal bovine serum (Sigma-Aldrich), 0.1% gentamicin (Thermo Fisher Scientific).

Techniques: Fluorescence

Journal: ACS Applied Bio Materials

Article Title: Real-Time Monitoring of the Cytotoxic Effect of Oxygen-Sensitive Fluorescent Poly(styrene-maleic anhydride) Nanoparticles Using Electrical-Substrate Impedance Sensing

doi: 10.1021/acsabm.5c01443

Figure Lengend Snippet: SEM images showing the cell–NP interaction after 7 days of culture with fluorescent SMA NPs. (Top panel) NIH 3T3 fibroblasts. (Bottom panel) bEnd3 brain endothelial cells. Turquoise hexagons point to SMA NPs attached to cells and intercalated into the extracellular matrix (ECM). The left and right panels are replicate samples. Scale bar = 10 μm. Zoomed micrograph scale bar = 400 nm.

Article Snippet: bEnd3 murine brain endothelial cells, obtained from the American Type Culture Collection (ATCC, CRL-2299, VA), were cultured in Gibco DMEM (low glucose) supplemented with 10% fetal bovine serum (Sigma-Aldrich), 0.1% gentamicin (Thermo Fisher Scientific).

Techniques:

Journal: Frontiers in Immunology

Article Title: Serum EMMPRIN/CD147 promotes the lung pre-metastatic niche in a D2A1 mammary carcinoma mouse model

doi: 10.3389/fimmu.2025.1568578

Figure Lengend Snippet: EMMPRIN promotes angiogenesis in the lung PMN. Mice were injected with the different tumor cells and treatments as described in the legend of Figure 1 . (A) Representative images of the endothelial cell marker CD31 staining in the lungs, and (C) their quantitation (n=9 for the D2A1-WT group, n=8 for the D2A1-KD group, n=7 for the healthy group, n=7 for the D2A1-WT + m161-pAb group, n=9 for the Healthy + rec. EMMRPIN group). Bar size is 100 μm. (B) Representative images of bEND3 endothelial cell migration length after their incubation for 18 h with serum-starvation media containing 5µg of lung lysates from the different experimental groups. (D) Quantitation of the migration length (n=14 for the D2A1-WT group, n=10 for the D2A1-KD group, n=8 for the healthy group, n=9 for the D2A1-WT + m161-pAb group, n=7 for the Healthy + rec. EMMRPIN group). Bar size is 150 μm. Data are presented as mean ± SEM. Three groups were analyzed using one-way ANOVA followed by Bonferroni’s post-hoc test, and two groups were compared using the non-parametric two-tailed Mann-Whitney t test. The increased CD31 staining of endothelial cells in the mice implanted with the D2A1-WT cells and their longer migration distance suggests that they proliferate and migrate, two properties necessary for angiogenesis, more than the endothelial cells in healthy mice or mice implanted with the D2A1-KD cells. The importance of EMMPRIN to this process is exemplified by the results from the addition of h161-pAb or the injection of recombinant EMMPRIN.

Article Snippet: The mouse brain endothelial cell line bEND3 (ATCC CRL-2299) was cultured in the same full medium that contained 1% glutamine, whereas D2A1 full medium contained 2% glutamine.

Techniques: Injection, Marker, Staining, Quantitation Assay, Migration, Incubation, Two Tailed Test, MANN-WHITNEY, Recombinant

Journal: Journal of Lipid Research

Article Title: Phosphatidylcholine synthesis and remodeling in brain endothelial cells

doi: 10.1016/j.jlr.2025.100773

Figure Lengend Snippet: Tracing of glycerophosphocholine metabolism. (A) Cellular acylation of lyso-phosphatidylcholine (LPC) by fatty acids (FA, or other acyl donors) yields phosphatidylcholine (PC) and upon de-acylation LPC is re-produced. This Lands cycle allows for side-chain exchange. (B) A library of synthetic head group-labeled (magenta) alkyne tracers featuring saturated (SaFA), mono-unsaturated (MUFA), or poly-unsaturated (PUFA) fatty acid side chains at the sn - 1 position were synthesized. The N-propargyl- sn 2 -lysophosphatidylcholine (LpPC) tracer family includes members with ω−6 (orange) and ω−3 (red) PUFA side chains. Also an ether derivative (LpPCO) containing an sn 1 -O-alkyl side chain (green) was synthesized. (C) bEND3 cells were incubated for 24 h with 50 μM LpPC containing the indicated side chain. Total lipids from cells were isolated, click-reacted and analyzed by TLC. Fluorescent bands were identified by co-migrating standards (closed arrowheads). Cellular pPC metabolites were detected as multiple bands. The double band migrating between LpPC and pPC putatively represents pSM (open arrowhead). ori, origin of application.

Article Snippet: The brain endothelial cell line bEND3 ( ) was obtained from ATCC (CRL-2299) and maintained in DMEM medium (Gibco, 31966021) containing 10% fetal calf serum (Gibco, A5256701) and 1% penicillin/streptomycin (Gibco, 15070063).

Techniques: Produced, Labeling, Synthesized, Incubation, Isolation

Journal: Journal of Lipid Research

Article Title: Phosphatidylcholine synthesis and remodeling in brain endothelial cells

doi: 10.1016/j.jlr.2025.100773

Figure Lengend Snippet: MS analysis of pPC metabolism in bEND3 cells over time. Cells were incubated with one of four LpPC tracers (50 μM) for various times. Total cellular lipids and internal standards were co-isolated and click-reacted before multiplexed MS analysis. All labeled pPC species were quantified (see supplemental Tables S1–S4 ) and the total cellular pPC content per 100,000 cells is plotted over time (A). Error bars smaller than symbol size are omitted. Cells incubated with (B) LpPC 16:0, (C) LpPC 18:2, (D) LpPC 20:4, or (E) LpPC 22:6 showed distinct distributions of labeled pPC species at each time point. Upon 24 h (magenta bars) of constant tracer supply and ongoing cellular lipid remodeling the labeled lipidome differed substantially from earlier time points. The species distribution of unlabeled control samples (green bars) is also shown. Labeled pPC species are presented as the molar fraction (mol%) of all labeled pPC species. N = 3.

Article Snippet: The brain endothelial cell line bEND3 ( ) was obtained from ATCC (CRL-2299) and maintained in DMEM medium (Gibco, 31966021) containing 10% fetal calf serum (Gibco, A5256701) and 1% penicillin/streptomycin (Gibco, 15070063).

Techniques: Incubation, Isolation, Labeling, Control

Journal: Journal of Lipid Research

Article Title: Phosphatidylcholine synthesis and remodeling in brain endothelial cells

doi: 10.1016/j.jlr.2025.100773

Figure Lengend Snippet: Analysis of LPC acylation and PC side chain remodeling in bEND cells. (A) Experimental scheme: For 30 min bEND3 cells were pulse-incubated with 50 μM LpPC containing the indicated side chain and 50 μM of the homologous isotope-labeled FA (red) to generated isotope-labeled symmetric pPC metabolites. Symmetric pPC lacking an isotope label was produced if the unlabeled homologous FA (magenta) served as acylation partner. Cells were chased in the presence of isotope-labeled oleate (green) for the indicated time during which side chain replacement yielded labeled asymmetric pPC. (B–E) The amount of selected pPC species is presented over time as detected by MS in lipid extracts from cells incubated with (B) LpPC 16:0, (C) LpPC 18:2, (D) LpPC 20:4 and (E) LpPC 22:6. The isotope-labeled symmetric pPC (red) and the symmetric pPC lacking an isotope label (magenta) were produced first during the pulse. Generated upon side chain remodeling, the pPC species carrying the side chain of the respective input LpPC as well as the 13C-oleate added during chase is highlighted in green. The sum of all pPC species labeled by 13C-oleate during the chase is shown in cyan. N = 3.

Article Snippet: The brain endothelial cell line bEND3 ( ) was obtained from ATCC (CRL-2299) and maintained in DMEM medium (Gibco, 31966021) containing 10% fetal calf serum (Gibco, A5256701) and 1% penicillin/streptomycin (Gibco, 15070063).

Techniques: Incubation, Labeling, Generated, Produced