Journal: Materials Today Bio

Article Title: Injectable versatile liquid-solid transformation implants alliance checkpoint blockade for magnetothermal dynamic-immunotherapy

doi: 10.1016/j.mtbio.2022.100442

Figure Lengend Snippet: Magnetothermal performance and therapeutic efficacy of F/A@P with various components in vitro. (a) Magnetothermal curves of different volumes and mass fractions of Fe 3 O 4 in F/A@P under AMF actuation and (b) the corresponding infrared thermal images. (c) Temperature curves of F/A@P over seven on-off cycles under AMF actuation. (d–e) Viability of 4T1 cells after different treatments assessed by CCK8 assay. (f) Live (green)/dead (red) fluorescence images of 4T1 cells after different treatments (scale bars: 50 ​μm). (g) FCM was used to determine the apoptosis level of 4T1 cells after various treatments. The data are shown as the mean ​± ​SD, n ​= ​3 per group; ∗p ​< ​0.05, ∗∗p ​< ​0.01, ∗∗∗p ​< ​0.001.

Article Snippet: 4T1 breast tumor cells were purchased from Procell (Wuhan, China), cultured in complete DMEM and grown in an atmosphere composed of 5% carbon dioxide (CO 2 ) and 95% air at 37 °C.

Techniques: Drug discovery, In Vitro, CCK-8 Assay, Fluorescence

Journal: Materials Today Bio

Article Title: Injectable versatile liquid-solid transformation implants alliance checkpoint blockade for magnetothermal dynamic-immunotherapy

doi: 10.1016/j.mtbio.2022.100442

Figure Lengend Snippet: Oxygen-irrelevant free radical-generating ability of F/A@P and the mechanism of ER stress in vitro. (a) UV–vis spectrum of oxidized TMB in different groups after AMF actuation for 400 ​s (inset: digital photos of the TMB solutions). (b) UV–vis spectrum of oxidized TMB after treatment with F/A@P under different AMF actuation times (inset: digital photos of the TMB solutions). (c) Absorbance curves of oxidized TMB based on a) and b). (d) Typical DCF fluorescence images of ROS levels in 4T1 cells after various treatments (scale bars: 50 ​μm). (e) Intracellular ROS levels after different treatments analyzed by FCM and (f) the corresponding quantitative analyses of ROS levels. (g) Intracellular GSH levels after different treatments. (h) Representative immunoblot results of various protein expression levels from each group. (i) Corresponding quantitative analyses of the ratios of p-PERK/GAPDH, ATF4/GAPDH and CHOP/GAPDH based on h). (The data are shown as the mean ​± ​SD, n ​= ​3 per group; n.s. represents no significance, ∗p ​< ​0.05, ∗∗p ​< ​0.01, ∗∗∗p ​< ​0.001 in comparison with the control group, ##p ​< ​0.01 in comparison with the ICG ​+ ​laser group).

Article Snippet: 4T1 breast tumor cells were purchased from Procell (Wuhan, China), cultured in complete DMEM and grown in an atmosphere composed of 5% carbon dioxide (CO 2 ) and 95% air at 37 °C.

Techniques: In Vitro, Fluorescence, Western Blot, Expressing, Comparison, Control

Journal: Materials Today Bio

Article Title: Injectable versatile liquid-solid transformation implants alliance checkpoint blockade for magnetothermal dynamic-immunotherapy

doi: 10.1016/j.mtbio.2022.100442

Figure Lengend Snippet: F/A@P mediates immunogenic cell death and DC maturation in vitro. (a) Schematic diagram of the coculture process of immature DCs with 4T1 cell residues in vitro. (b) Immunofluorescence images of HMGB1, CRT, HSP70, and HSP90 expression in 4T1 cells (scale bars: 25 ​μm). Nuclei: blue, DAPI-labeled; specific cell surface proteins: green, FITC-IgG secondary antibody-stained. (c) The expression of CD80 and CD86 on the surface of the DCs in each group was analyzed by FCM. (d) Corresponding quantitative analysis of the DC maturation rates and levels of the cytokines IL-6, IL-12 and TNF-α in the supernatants of the DC culture medium. (The data are shown as the mean ​± ​SD, n ​= ​3 per group; n.s. represents no significance, ∗p ​< ​0.05, ∗∗p ​< ​0.01, ∗∗∗p ​< ​0.001 in comparison with the control group).

Article Snippet: 4T1 breast tumor cells were purchased from Procell (Wuhan, China), cultured in complete DMEM and grown in an atmosphere composed of 5% carbon dioxide (CO 2 ) and 95% air at 37 °C.

Techniques: In Vitro, Immunofluorescence, Expressing, Labeling, Staining, Comparison, Control

Journal: Materials Today Bio

Article Title: Injectable versatile liquid-solid transformation implants alliance checkpoint blockade for magnetothermal dynamic-immunotherapy

doi: 10.1016/j.mtbio.2022.100442

Figure Lengend Snippet: Antitumor effect of F/A@P-mediated MDT combined with CTLA4 blockade therapy in an orthotopic 4T1 bilateral tumor-bearing mouse model. (a) Schematic diagram of F/A@P-mediated MHT combined with CTLA4 blockade therapy to inhibit primary and distant tumors. (b) Digital photographs of the mice in the five groups at 0, 1, 7, 14 and 21 days after various treatments. (c) Primary tumor growth curves and the average primary tumor growth curve of the mice in each group (n ​= ​6). (d) Distant tumor growth curves and average distant growth curve in each group (n ​= ​6). (e) Primary tumor inhibition rates and (f) distant tumor inhibition rates of the mice in each group. (g) Body weights and (h) morbidity-free survival of the mice in each group. (i) Digital photographs and H&E staining images of representative pulmonary metastatic nodules (circles) from the mice in each group at the end of treatment (scale bars: 100 ​μm). (The data are shown as the mean ​± ​SD, n ​= ​6 per group; n.s. represents no significance, ∗p ​< ​0.05, ∗∗p ​< ​0.01, ∗∗∗p ​< ​0.001 in comparison with the control group, #p ​< ​0.05, ##p ​< ​0.01, ###p ​< ​0.001 in comparison with the F/A@P ​+ ​AMF group).

Article Snippet: 4T1 breast tumor cells were purchased from Procell (Wuhan, China), cultured in complete DMEM and grown in an atmosphere composed of 5% carbon dioxide (CO 2 ) and 95% air at 37 °C.

Techniques: Inhibition, Staining, Comparison, Control

Journal: Materials Today Bio

Article Title: Injectable versatile liquid-solid transformation implants alliance checkpoint blockade for magnetothermal dynamic-immunotherapy

doi: 10.1016/j.mtbio.2022.100442

Figure Lengend Snippet: Immune responses to combination therapy in an orthotopic bilateral 4T1 tumor-bearing mouse model. (a) FCM diagrams and quantification analysis of DC maturation in drain LNs adjacent to the primary tumors. (b) FCM diagrams and quantification analysis of DC maturation in spleens. (c) FCM diagrams and (d) quantification analysis of CTLs and Tregs in spleens. (e) FCM diagrams and (f) quantification analysis of CTLs and Tregs in distant tumors. (g) FCM diagrams and (h) quantification analysis of T EM cells in the mouse spleens of the control group and F/A@P ​+ ​AMF ​+ ​anti-CTLA4 group. (i) Proportions of Teff cells in secondary tumors based on (e) and (f). (j) Teff:Treg and CTL:Treg ratios in the secondary tumors. (The data are shown as the mean ​± ​SD, n ​= ​3 per group; n.s. represents no significance, ∗p ​< ​0.05, ∗∗p ​< ​0.01, ∗∗∗p ​< ​0.001 in comparison with the control group, #p ​< ​0.05, ##p ​< ​0.01, ###p ​< ​0.001 in comparison with the F/A@P ​+ ​AMF group).

Article Snippet: 4T1 breast tumor cells were purchased from Procell (Wuhan, China), cultured in complete DMEM and grown in an atmosphere composed of 5% carbon dioxide (CO 2 ) and 95% air at 37 °C.

Techniques: Control, Comparison

Journal: Cancer cell

Article Title: Fasting mimicking diet reduces HO-1 to promote T cell-mediated tumor cytotoxicity

doi: 10.1016/j.ccell.2016.06.005

Figure Lengend Snippet: (A-D) 12-week old female BALB/c mice were grafted with breast cancer (4T1) cells and subject to (A) multiple cycles of FMD or STS alone or to (C, D) a combination of FMD and the chemotherapy drugs (C) doxorubicin (DXR) or (D) cyclophosphamide (CP). (B) Circulating IGF-1 levels at the end of STS or FMD in (A) were measured. (E) 12-week old female C57BL/6 mice were grafted with melanoma (B16) cells and subject to multiple cycles of FMD alone or in combination with DXR. Tumor volume at multiple time-points (on the left) and immediately prior to euthanasia (on the right) are reported for each set of treatments. The animals receiving chemotherapy were injected at the end of each FMD cycle (shaded area). [(A, B) n=10, (C) n=13, (D) n=10, and (E) n=15]. Data represented as mean ± SEM. One-way ANOVA (Tukey post-analysis test) was performed. p-values <0.05, 0.01 and 0.001 are indicated as *, **, and ***, respectively. See also Figure S1.

Article Snippet: Cancer cell lines and tumor cell injection MCF7 human breast adenocarcinoma (gift from Amy Lee – University of Southern California), 4T1- luc murine breast cancer (SibTech, Inc.) and B16 murine melanoma (gift from Noah Craft, UCLA) cell lines were cultured in high glucose (4.5g/L) Dulbecco's modified Eagle's medium (DMEM) supplemented with 10% FBS (Thermo Fisher Scientific Inc.) at 37°C and 5% CO 2 .

Techniques: Injection

Journal: Cancer cell

Article Title: Fasting mimicking diet reduces HO-1 to promote T cell-mediated tumor cytotoxicity

doi: 10.1016/j.ccell.2016.06.005

Figure Lengend Snippet: (A) Bone marrow collected from BALB/c mice undergoing FMD/DXR treatments (see Figure S3A) was collected at the end of the experiment and analyzed with FACS (n=6) to assess the amount of Common Lymphoid Progenitors (CLP). (B) Breast cancer (4T1) tumor tissues collected at the end of the experiment (see Figure S3A) were analyzed using immunohistochemistry to assess CD3+/CD8+ TILs (n=8) (quantification on the right). (C, E) CD3+/CD8+ and (D, E) CD3+/CD4+/CD25+were also assessed by FACS analysis (n=7) in tumor tissue collected from a separate, equivalent experiment (see Figure 1C). (F) Melanoma (B16) tissue collected (see Figure 1E) was also processed to assess the levels of TILs (quantification on the right). Bar scale for DAPI, CD3 and CD8 is 75 μm. Bar scale for CD3+/CD8+ is 12.5 μm Data represented as mean ± SEM. The significance of the differences between experimental groups was determined by using one-way ANOVA (Tukey post-analysis test). p-values <0.05, 0.01 and 0.001 are indicated as *, **, and ***, respectively. See also Figure S2.

Article Snippet: Cancer cell lines and tumor cell injection MCF7 human breast adenocarcinoma (gift from Amy Lee – University of Southern California), 4T1- luc murine breast cancer (SibTech, Inc.) and B16 murine melanoma (gift from Noah Craft, UCLA) cell lines were cultured in high glucose (4.5g/L) Dulbecco's modified Eagle's medium (DMEM) supplemented with 10% FBS (Thermo Fisher Scientific Inc.) at 37°C and 5% CO 2 .

Techniques: Immunohistochemistry

Journal: Cancer cell

Article Title: Fasting mimicking diet reduces HO-1 to promote T cell-mediated tumor cytotoxicity

doi: 10.1016/j.ccell.2016.06.005

Figure Lengend Snippet: (A) The effect of FMD on breast tumor growth (4T1) in immunodeficient BALB/c nude mice (n=15) (see Figure S4A-E); (B) The survival of wild type (WT) and nude mice injected with high dose of DXR and undergoing ad lib or FMD regimen. (C-H) 4T1 breast tumor-bearing mice were treated with DXR, FMD+DXR alone or in combination with [.alpha]CD8 monoclonal antibody and (C-E) circulating levels of (C, E) CD3+/CD8+ and (D, E) CD3+/CD4+/CD25+ were determined by FACS (n=7). (F) Tumor volumes of FMD+DXR and FMD+DXR+αCD8 were measured at multiple time points, and (G) TILs were also assessed in tumor samples collected from the same animals. (H) Circulating lymphocytes from DXR, FMD+DXR, DXR+αCD8, and FMD+DXR+αCD8 these animals were collected and cultured ex vivo with 4T1 cells for 24 hours and viability was assessed by MTT reduction. (I) Mice were immunized by subcutaneous inoculation (in the left flank) with 4T1 breast cancer cells preconditioned in vitro with either ad lib- (2 g/L glucose+10% FBS) or STS-medium (0.5 g/L glucose+1% FBS) with or without doxorubicin (5 μM). 7 days after the immunization, the same animals were inoculated with naïve 4T1 cells in the right flank (n=10). (J, K) Tumor progression for the immunization (left) and the naïve (right) sides are reported. Data represented as mean ± SEM. One-way ANOVA (Tukey post-analysis test) and Log-rank (Mantel-Cox) test (survival) was performed. Comparisons between groups were performed with Student's t test. p-values <0.05, 0.01 and 0.001 are indicated as *, **, and ***, respectively. See also Figure S3.

Article Snippet: Cancer cell lines and tumor cell injection MCF7 human breast adenocarcinoma (gift from Amy Lee – University of Southern California), 4T1- luc murine breast cancer (SibTech, Inc.) and B16 murine melanoma (gift from Noah Craft, UCLA) cell lines were cultured in high glucose (4.5g/L) Dulbecco's modified Eagle's medium (DMEM) supplemented with 10% FBS (Thermo Fisher Scientific Inc.) at 37°C and 5% CO 2 .

Techniques: Injection, Cell Culture, Ex Vivo, In Vitro

Journal: Cancer cell

Article Title: Fasting mimicking diet reduces HO-1 to promote T cell-mediated tumor cytotoxicity

doi: 10.1016/j.ccell.2016.06.005

Figure Lengend Snippet: (A) HO-1 expression levels of grafted 4T1 tumor and normal (liver and skeletal muscle) tissues collected from ad lib fed and mice undergoing STS or FMD regimen were analyzed with qRT-PCR. (B) HO-1 levels in 4T1 cells following 48 hours of in vitro STS were measured by Western blotting (n=3) (Blot was captured with Bio-Rad ChemiDoc, and unedited, representative bands are shown. (C, E, G) Viability of 4T1 cells was determined by MTT reduction following (C) DXR and hemin (10 M), (E) CP and hemin (10 M), and (G) CP and ZnPP (20 M). (D, F) Viability of 4T1 cell stably over-expressing HO-1 (pHO-1) or empty vector (pEV) was determined by MTT following (D) DXR and (F) CP under control and STS conditions. Data represented as mean ± SEM. The significance of the differences between experimental groups was determined by using one-way ANOVA (Tukey post-analysis test). Comparisons between groups were performed with Student's t test. p-values <0.05, 0.01 and 0.001 are indicated as *, **, and ***, respectively. See also Figure S5.

Article Snippet: Cancer cell lines and tumor cell injection MCF7 human breast adenocarcinoma (gift from Amy Lee – University of Southern California), 4T1- luc murine breast cancer (SibTech, Inc.) and B16 murine melanoma (gift from Noah Craft, UCLA) cell lines were cultured in high glucose (4.5g/L) Dulbecco's modified Eagle's medium (DMEM) supplemented with 10% FBS (Thermo Fisher Scientific Inc.) at 37°C and 5% CO 2 .

Techniques: Expressing, Quantitative RT-PCR, In Vitro, Western Blot, Stable Transfection, Plasmid Preparation, Control

Journal: Cancer cell

Article Title: Fasting mimicking diet reduces HO-1 to promote T cell-mediated tumor cytotoxicity

doi: 10.1016/j.ccell.2016.06.005

Figure Lengend Snippet: (A-C) 4T1 tumor-bearing mice were treated with (A) ZnPP (40 mg/kg/day; IP; n=7), (B) STS and hemin (30 mg/kg/day; IP; n=7), and (C) FMD and hemin and DXR. (D) Mice bearing 4T1 tumors stably over-expressing HO-1 (pHO-1), or empty vector (pEV) were treated with DXR under ad lib or FMD regimens (n=10-15) (see Figure 1C). Data represented as mean ± SEM. One-way ANOVA (Tukey post-analysis test) was performed. p-values <0.05, 0.01 and 0.001 are indicated as *, **, and ***, respectively. See also Figure S6.

Article Snippet: Cancer cell lines and tumor cell injection MCF7 human breast adenocarcinoma (gift from Amy Lee – University of Southern California), 4T1- luc murine breast cancer (SibTech, Inc.) and B16 murine melanoma (gift from Noah Craft, UCLA) cell lines were cultured in high glucose (4.5g/L) Dulbecco's modified Eagle's medium (DMEM) supplemented with 10% FBS (Thermo Fisher Scientific Inc.) at 37°C and 5% CO 2 .

Techniques: Stable Transfection, Expressing, Plasmid Preparation

Journal: Cancer cell

Article Title: Fasting mimicking diet reduces HO-1 to promote T cell-mediated tumor cytotoxicity

doi: 10.1016/j.ccell.2016.06.005

Figure Lengend Snippet: Mice bearing 4T1 or 4T1-pHO-1 breast tumors under FMD were treated with DXR and either hemin (30 mg/kg/day; IP) or CD8+ specific neutralizing monoclonal antibodies (αCD8; clone YTS 169.4) (n=13-15) (see Figures 3F and ​and5D).5D). (A) Tumor volumes were measured at multiple time-points (left) and immediately prior to euthanasia (right). (B-D) CD3+/CD8+ (CTL) and CD3+/CD4+/CD25+ (Treg)lymphocytes from the tumor bed was analyzed by FACS. Data represented as mean ± SEM. One-way ANOVA (Tukey post-analysis test) was performed. p-values <0.05, 0.01 and 0.001 are indicated as *, **, and ***, respectively.

Article Snippet: Cancer cell lines and tumor cell injection MCF7 human breast adenocarcinoma (gift from Amy Lee – University of Southern California), 4T1- luc murine breast cancer (SibTech, Inc.) and B16 murine melanoma (gift from Noah Craft, UCLA) cell lines were cultured in high glucose (4.5g/L) Dulbecco's modified Eagle's medium (DMEM) supplemented with 10% FBS (Thermo Fisher Scientific Inc.) at 37°C and 5% CO 2 .

Techniques: Bioprocessing

Journal: Cells

Article Title: Interaction between Chromodomain Y-like Protein and Androgen Receptor Signaling in Sertoli Cells Accounts for Spermatogenesis

doi: 10.3390/cells13100851

Figure Lengend Snippet: AR is colocalized and interacts with CDYL in male testes and Sertoli cells. ( A ) Immunohistochemical staining of AR and CDYL in testes obtained from wild-type and ARKO mice. Bar = 50 um. ( B ) Localization of AR and CDYL (upper) and co-localization of AR, CDYL, and DAPI (bottom) in the testes obtained from wild-type mice by immunofluorescence analysis. Bar = 20 μm. ( C ) Interaction was observed between AR and CDYL in wild-type mouse testes. IgG was used as the control for Western blotting in each group. “Input” means the sample on 10% of volume used for IP. ( D ) Protein expression patterns of AR and CDYL in wild-type mouse testes and ARKO mice by Western blotting. GAPDH was used as the internal control. ( E ) AR and CDYL mRNA expressions were detected in testicular tissues between wild-type and ARKO mice by quantitative RT–PCR assay. ( n ≥ 3) * p ˂ 0.05, by unpaired two-tailed Student t tests was significant compared with the control. Data are expressed as the mean ± standard error of three samples per group. S: Sertoli cell; L: Leydig cell; M: myoid cell; SP: spermatids.

Article Snippet: The TM4 mouse Sertoli cell line (BCRC-60254) was procured from the Bioresource Collection and Research Center (BCRC) in Hsinchu, Taiwan.

Techniques: Immunohistochemical staining, Staining, Immunofluorescence, Control, Western Blot, Expressing, Quantitative RT-PCR, Two Tailed Test

Journal: Cells

Article Title: Interaction between Chromodomain Y-like Protein and Androgen Receptor Signaling in Sertoli Cells Accounts for Spermatogenesis

doi: 10.3390/cells13100851

Figure Lengend Snippet: AR and CDYL regulate downstream AR-targeted genes. ChIP and luciferase assays demonstrate the CDYL ( A ) or TNP1 ( B ) genes in the presence of AR binding sequences. Schematic of a putative binding site for the transcription factor, androgen-responsive elements (ARE) sequences on the CDYL or TNP1 promoter. Promoter occupancy of AR on CDYL or TNP1 promoter by ChIP. CDYL or TNP1 upstream regions starting at position −3000 were introduced into the pGL3-Basic plasmid before the luciferase reporter gene. The siRNA-mediated knockdown ( C ) or overexpression ( D ) of AR or CDYL substantially changed the CDYL or TNP1 promoter activity in TM4 cells through dual-luciferase assays. * p ˂ 0.05 for one-way ANOVA was significant compared with the control. ( n ≥ 3) in each group, and error bars represent ± SD.

Article Snippet: The TM4 mouse Sertoli cell line (BCRC-60254) was procured from the Bioresource Collection and Research Center (BCRC) in Hsinchu, Taiwan.

Techniques: Luciferase, Binding Assay, Plasmid Preparation, Knockdown, Over Expression, Activity Assay, Control

Journal: Cells

Article Title: Interaction between Chromodomain Y-like Protein and Androgen Receptor Signaling in Sertoli Cells Accounts for Spermatogenesis

doi: 10.3390/cells13100851

Figure Lengend Snippet: mRNA expression of AR , CDYL , and TNP1 association by CDYL rescue in Sertoli cells. mRNA expression of AR , CDYL , and TNP1 association among control, AR siRNA-treated, and CDYL siRNA-treated groups in TM4 cells by non-treat and CDYL rescue. mRNA expression of AR ( A , B ), CDYL ( C , D ), or TNP1 ( E , F ) were detected after being transiently transfected with control, AR , or CDYL siRNA-treated ( A , C , E ). Moreover, these three groups were treated with 200 ng CDYL recombinant protein ( B , D , F ) for 24 h in TM4 cells. * p ˂ 0.05 for one-way ANOVA was significant compared with the control. ( n ≥ 3) in each group, and error bars represent ± SD.

Article Snippet: The TM4 mouse Sertoli cell line (BCRC-60254) was procured from the Bioresource Collection and Research Center (BCRC) in Hsinchu, Taiwan.

Techniques: Expressing, Control, Transfection, Recombinant

Journal: Cells

Article Title: Interaction between Chromodomain Y-like Protein and Androgen Receptor Signaling in Sertoli Cells Accounts for Spermatogenesis

doi: 10.3390/cells13100851

Figure Lengend Snippet: Decreased expression of CDYL in the human testes from patients with azoospermia . Immunohistochemistry indicated the expression of AR and CDYL protein in testicular histology from obstructive azoospermia (active spermatogenesis) and non-obstructive azoospermia (defective spermatogenesis), including Sertoli cell-only syndrome (SCOS) and maturation arrest (halted at the primary spermatocyte stage). Bar = 50 μm. The AR and CDYL signals in patients with normal group ( A , D ), Sertoli cell-only syndrome ( B , E ), and maturation arrest ( C , F ) ( n = 1 patient in every group). S: Sertoli cell; L: Leydig cell; M: myoid cell; SP: spermatids.

Article Snippet: The TM4 mouse Sertoli cell line (BCRC-60254) was procured from the Bioresource Collection and Research Center (BCRC) in Hsinchu, Taiwan.

Techniques: Expressing, Immunohistochemistry

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Combining photodynamic therapy and cascade chemotherapy for enhanced tumor cytotoxicity: the role of CTT 2 P@B nanoparticles

doi: 10.3389/fbioe.2024.1361966

Figure Lengend Snippet: ROS generation detection in vitro . (A) The release of singlet oxygen from each drug was detected in vitro under dark conditions. Data were presented as the mean ± SD ( n = 3). (B) Detection of singlet oxygen release of each drug under 660 nm laser irradiation at a power density of 280 mW cm -2 was performed in vitro . Data were presented as the mean ± SD ( n = 3). (C) ROS generation was observed in 4T1 cells treated with each medication while being exposed to laser irradiation at a wavelength of 660 nm and intensity of 280 mW cm −2 . Bright: Bright field. DCF: Green fluorescence represents intracellular ROS. Merge: Superimpose the image. Scale Bar = 100 μm.

Article Snippet: The murine breast cancer cells (4T1) were procured from Icell Biotech Co., Ltd. (Shanghai, China).

Techniques: In Vitro, Irradiation, Fluorescence

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Combining photodynamic therapy and cascade chemotherapy for enhanced tumor cytotoxicity: the role of CTT 2 P@B nanoparticles

doi: 10.3389/fbioe.2024.1361966

Figure Lengend Snippet: Evaluation of mitochondrial targeting function. (A) Confocal images of the mitochondrial sites of TPPOH 2 , CTT 2 , CTT 2 P, and CTT 2 P@B NPs in 4T1 cells. Mito-Tracker Green was used to stain the mitochondria in the green channel. The red channel was derived from the emission of the photosensitizer fraction (PS) itself. Merge stands for superimposed image. The Green and red curves in the Plot Profile represent the gray value of Mito-Tracker Green and PS, respectively. Scale bar = 20 μm. (B) Flow cytometry JC-1 method was used to analyze the mitochondrial function of cells treated with different drugs. Red fluorescence: normal mitochondria (J-aggregate); Green fluorescence: depolarized mitochondria (J-monomer).

Article Snippet: The murine breast cancer cells (4T1) were procured from Icell Biotech Co., Ltd. (Shanghai, China).

Techniques: Staining, Derivative Assay, Flow Cytometry, Fluorescence

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Combining photodynamic therapy and cascade chemotherapy for enhanced tumor cytotoxicity: the role of CTT 2 P@B nanoparticles

doi: 10.3389/fbioe.2024.1361966

Figure Lengend Snippet: Evaluation of cell death in vitro through cytotoxicity assessment and examination of apoptosis and necrosis. (A) The in vitro cytotoxicity of 4T1 cells treated with CPT, TPPOH 2 , CTT 2 , CTT 2 P, and CTT 2 P@B NPs in the dark was assessed using the CCK-8 assay. Data were presented as the mean ± SD ( n = 5). ** p < 0.01, **** p < 0.0001. (B) The in vitro cytotoxicity of 4T1 cells treated with TPPOH 2 , CTT 2 , CTT 2 P and CTT 2 P@B NPs under laser irradiation (660 nm, 280 mW cm −2 ) was determined using the CCK-8 assay. Data were presented as the mean ± SD ( n = 5). ** p < 0.01, **** p < 0.0001. (C) Cell apoptosis and necrosis were analyzed using flow cytometry with Annexin V-FITC/PI double staining following treatment with various drugs at a concentration of 5 μM.

Article Snippet: The murine breast cancer cells (4T1) were procured from Icell Biotech Co., Ltd. (Shanghai, China).

Techniques: In Vitro, CCK-8 Assay, Irradiation, Flow Cytometry, Double Staining, Concentration Assay

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Combining photodynamic therapy and cascade chemotherapy for enhanced tumor cytotoxicity: the role of CTT 2 P@B nanoparticles

doi: 10.3389/fbioe.2024.1361966

Figure Lengend Snippet: Biodistribution in vivo . (A) Blood compatibility test of CPT, TPPOH 2 , CTT 2 , CTT 2 P, CTT 2 P@B NPs. Data were presented as the mean ± SD ( n = 3). *** p < 0.001, **** p < 0.0001. (B) Time-lapse live fluorescence imaging of mice with 4T1 tumors following the administration of free CTT 2 P and CTT 2 P@B NPs via intravenous injection. (C) Fluorescent images of major organs and tumors were obtained 24 h after injection, using ex vivo methods. (D) The mean fluorescence intensity of each organ and tumor was used to determine the biodistribution of free CTT 2 P and CTT 2 P@B NPs in mice. Data were presented as the mean ± SD ( n = 3). * p < 0.05.

Article Snippet: The murine breast cancer cells (4T1) were procured from Icell Biotech Co., Ltd. (Shanghai, China).

Techniques: In Vivo, Fluorescence, Imaging, Injection, Ex Vivo

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Combining photodynamic therapy and cascade chemotherapy for enhanced tumor cytotoxicity: the role of CTT 2 P@B nanoparticles

doi: 10.3389/fbioe.2024.1361966

Figure Lengend Snippet: In vivo anti-tumor study of each drug in 4T1 tumor-bearing mice. (A) Tumor images of different drug administration treatments after the antitumor study. (B) During the administration, the growth of tumors in mice was observed in each group receiving treatment. Data were presented as the mean ± SD ( n = 5), ** p < 0.01. (C) The weight of mice in each treatment group was monitored throughout the administration period. Data were presented as the mean ± SD ( n = 5).

Article Snippet: The murine breast cancer cells (4T1) were procured from Icell Biotech Co., Ltd. (Shanghai, China).

Techniques: In Vivo

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Combining photodynamic therapy and cascade chemotherapy for enhanced tumor cytotoxicity: the role of CTT 2 P@B nanoparticles

doi: 10.3389/fbioe.2024.1361966

Figure Lengend Snippet: Investigation of the effects of each medication on mice with 4T1 tumors through pathological examination. (A) After administering various medications, the major organs (including the heart, liver, spleen, lung, and kidney) were subjected to H&E staining. Scale bar = 50 μm. (B) After administering various medications, tumors were subjected to H&E staining and TUNEL staining. Scale bar = 50 μm.

Article Snippet: The murine breast cancer cells (4T1) were procured from Icell Biotech Co., Ltd. (Shanghai, China).

Techniques: Medications, Staining, TUNEL Assay

Journal: Scientific Reports

Article Title: Fast and efficient isolation of murine circulating tumor cells using screencell technology for pre-clinical analyzes

doi: 10.1038/s41598-024-66032-x

Figure Lengend Snippet: Different volumes of WT mice blood collected either within ( A ) heparin tubes or ( B ) EDTA tubes were filtered with ScreenCell Cyto kits. Similar volumes of PyMT cancer mice blood were also processed with ScreenCell Cyto kits ( C ). To evaluate the correct morphology of cancer cells in EDTA tubes, MCF7, and 4T1 breast cancer cell lines were spiked into WT mice blood and processed with ScreenCell Cyto kits. The images were taken using 40X magnification ( D ). To evaluate the recovery rate of ScreenCell technology, either 0, 5, or 10 MCF7 cells were spiked into 100 µl of blood samples of healthy WT mice. Captured cancer cells were counted on Cyto ISs and recovery rate was calculated in percentage. Data are presented as mean ± SEM (n = 6) ( E ). All experiments were repeated at least 2 times.

Article Snippet: 4T1 mouse breast cancer cell line (passages 12–15) and MCF7 human breast cancer cell line (passages 7–10) were received from Inserm U1197 (Paul Brousse Hospital, Villejuif, France) and cultured in Dulbecco’s modified Eagle medium (DMEM) (Gibco, Life Technologies, USA) supplemented with 10% FBS (Pan biotech, Aidenbach, Germany) and 1% Penicillin/Streptomycin (P/S) (Thermofisher Scientific, Waltham, MA, USA).

Techniques:

Journal: Scientific Reports

Article Title: Fast and efficient isolation of murine circulating tumor cells using screencell technology for pre-clinical analyzes

doi: 10.1038/s41598-024-66032-x

Figure Lengend Snippet: This figure represents the 4T1 cancer cells isolated using ScreenCell Cyto kits either immediately after the blood draw (left image, 40X) or after 24 h of preservation at 4 °C in EDTA tubes (right image, 40X). This experiment was performed 4 times. T0: time zero.

Article Snippet: 4T1 mouse breast cancer cell line (passages 12–15) and MCF7 human breast cancer cell line (passages 7–10) were received from Inserm U1197 (Paul Brousse Hospital, Villejuif, France) and cultured in Dulbecco’s modified Eagle medium (DMEM) (Gibco, Life Technologies, USA) supplemented with 10% FBS (Pan biotech, Aidenbach, Germany) and 1% Penicillin/Streptomycin (P/S) (Thermofisher Scientific, Waltham, MA, USA).

Techniques: Isolation, Preserving

Journal: Scientific Reports

Article Title: Fast and efficient isolation of murine circulating tumor cells using screencell technology for pre-clinical analyzes

doi: 10.1038/s41598-024-66032-x

Figure Lengend Snippet: ScreenCell Cyto kits can isolate both single and cluster CTCs. ( A ) After spiking into WT mice blood, both single and cluster 4T1 cells were captured by ScreenCell Cyto kits. ( B ) Single and cluster CTCs were also efficiently captured from breast cancer mice blood. In all conditions, CTCs were distinguishable from normal epithelial cells according to their morphological features. These experiments were performed at least 3 times. All images are taken using 40X magnification, except for the CTC cluster (lower right) which is in 20X.

Article Snippet: 4T1 mouse breast cancer cell line (passages 12–15) and MCF7 human breast cancer cell line (passages 7–10) were received from Inserm U1197 (Paul Brousse Hospital, Villejuif, France) and cultured in Dulbecco’s modified Eagle medium (DMEM) (Gibco, Life Technologies, USA) supplemented with 10% FBS (Pan biotech, Aidenbach, Germany) and 1% Penicillin/Streptomycin (P/S) (Thermofisher Scientific, Waltham, MA, USA).

Techniques:

Journal: Scientific Reports

Article Title: Fast and efficient isolation of murine circulating tumor cells using screencell technology for pre-clinical analyzes

doi: 10.1038/s41598-024-66032-x

Figure Lengend Snippet: DNA concentration from WT mouse blood with or without spiked 4T1 cells isolated by ScreenCell MB device.

Article Snippet: 4T1 mouse breast cancer cell line (passages 12–15) and MCF7 human breast cancer cell line (passages 7–10) were received from Inserm U1197 (Paul Brousse Hospital, Villejuif, France) and cultured in Dulbecco’s modified Eagle medium (DMEM) (Gibco, Life Technologies, USA) supplemented with 10% FBS (Pan biotech, Aidenbach, Germany) and 1% Penicillin/Streptomycin (P/S) (Thermofisher Scientific, Waltham, MA, USA).

Techniques: Concentration Assay, Isolation

Journal: Scientific Reports

Article Title: Fast and efficient isolation of murine circulating tumor cells using screencell technology for pre-clinical analyzes

doi: 10.1038/s41598-024-66032-x

Figure Lengend Snippet: 4T1 cells were spiked into WT mice blood and filtered with ScreenCell MB kits. ISs were then directly inserted into 24 well plates and cultured for 10 days. The images of D0, D5, and D10 with RAL coloration are taken using 40X, and D10 without coloration is taken with 20X magnification. This experiment was performed 4 times. D: Day.

Article Snippet: 4T1 mouse breast cancer cell line (passages 12–15) and MCF7 human breast cancer cell line (passages 7–10) were received from Inserm U1197 (Paul Brousse Hospital, Villejuif, France) and cultured in Dulbecco’s modified Eagle medium (DMEM) (Gibco, Life Technologies, USA) supplemented with 10% FBS (Pan biotech, Aidenbach, Germany) and 1% Penicillin/Streptomycin (P/S) (Thermofisher Scientific, Waltham, MA, USA).

Techniques: Cell Culture

Journal: International Journal of Nanomedicine

Article Title: Improving the solubility and in vitro cytotoxicity (anticancer activity) of ferulic acid by loading it into cyclodextrin nanosponges

doi: 10.2147/IJN.S206350

Figure Lengend Snippet: MTT viability assay for ( A ) MCF7 and ( B ) 4T1 cell lines treated with different formulations of ferulic acid (FA)-loaded cyclodextrin nanosponges (NS) and various interval times at 570 nm.

Article Snippet: MCF7 (human breast cancer) and 4T1 (mouse breast cancer) cell lines were purchased from Pasteur Institute of Iran.

Techniques: MTT Viability Assay

Journal: International Journal of Nanomedicine

Article Title: Improving the solubility and in vitro cytotoxicity (anticancer activity) of ferulic acid by loading it into cyclodextrin nanosponges

doi: 10.2147/IJN.S206350

Figure Lengend Snippet: 4T1 cell lines treated with different formulations: control cells ( A ); ferulic acid, 250 µM ( B ); nanosponges, 250 µM ( C ); ferulic acid-loaded nanosponges, 250 µM ( D ). Microscopic magnification ×10 K, after 24 hrs of incubation.

Article Snippet: MCF7 (human breast cancer) and 4T1 (mouse breast cancer) cell lines were purchased from Pasteur Institute of Iran.

Techniques: Incubation

Journal: Molecular Pharmaceutics

Article Title: Non-Invasive Detection of Passively Targeted Poly(ethylene glycol) Nanocarriers in Tumors

doi: 10.1021/mp2003913

Figure Lengend Snippet: Passive tumor distribution of PEG nanocarriers measured non-invasively using a SkinSkan® spectrofluorometer. Nanocarriers (0.5 mM) were intravenously administered to 4T1 tumor-bearing balb/c mice and fluorescence spectra were collected from tumor and a contralateral skin (λexc.: 480 nm; λem.: 515 nm for 10–30 kDa and 520 nm for 40–60 kDa). Each column and error bar represents the mean±SD (n = 5–7). Individual comparisons between the groups were determined by Student’s t-test using the Microsoft Excel 2008. *: p<0.05, tumor vs. corresponding dermal control site.

Article Snippet: The 4T1 murine breast cancer cell line was a generous gift from Dr. Michael Reiss, The Cancer Institute of New Jersey (New Brunswick, NJ).

Techniques: Fluorescence, Control

Journal: Molecular Pharmaceutics

Article Title: Non-Invasive Detection of Passively Targeted Poly(ethylene glycol) Nanocarriers in Tumors

doi: 10.1021/mp2003913

Figure Lengend Snippet: (A) Non-invasive images showing the passive tumor distribution of PEG nanocarriers. The images were obtained on an IVIS® 100 imaging system using the excitation and emission filters corresponding to green fluorescent protein (GFP). The first two animals (balb/c mice with 4T1 tumor), starting from the left, were untreated (controls), whereas the remaining three animals were administered nanocarriers (0.5 mM) intravenously; (B) Regions of interest for tumor and control areas are shown in a mouse injected with 40 kDa nanocarrier for average efficient quantitation; and (C) Plots of average fluorescence from tumor and control site at skin’s surface against time. Each point represents the mean±SD (n = 3). Individual comparisons between groups were determined by Student’s t-test using the Microsoft Excel 2008. *: p<0.01, tumor vs. corresponding control site.

Article Snippet: The 4T1 murine breast cancer cell line was a generous gift from Dr. Michael Reiss, The Cancer Institute of New Jersey (New Brunswick, NJ).

Techniques: Imaging, Control, Injection, Quantitation Assay, Fluorescence

Journal: Molecular Pharmaceutics

Article Title: Non-Invasive Detection of Passively Targeted Poly(ethylene glycol) Nanocarriers in Tumors

doi: 10.1021/mp2003913

Figure Lengend Snippet: Ex vivo distribution studies: (A) Tissue distribution at 24 hrs; (B) Tumor distribution at 24 and 96 hrs; and (C) Plasma distribution at 24 and 96 hrs. PEG nanocarriers (0.5 mM) were intravenously administered to balb/c mice bearing 4T1 tumors and animals were euthanized to collect tissues. Each column and error bar represents the mean±SD (n = 5–7). The statistical analyses were carried out using GraphPad Prism v.4 as follows: (A) Two-way ANOVA and individual comparison between the groups were determined using Bonferroni posttests. The statistically significant groups (p<0.05, 60 kDa vs. other nanocarriers) are marked as * (10 kDa); & (20 kDa); # (30 kDa); and + (40 kDa); (B) One-way ANOVA and individual comparison between the groups were determined using Tukey posttests. The statistically significant groups (p<0.05, 60 kDa vs. other nanocarriers are marked as * (10, 20 or 30 kDa); and # (40 kDa at 96 hrs); (C) One-way ANOVA and comparison between the groups were determined using Tukey posttests. The statistically significant groups (p<0.05, 60 kDa vs. other nanocarriers) are denoted by * (10 or 20 kDa); & (30 kDa); # (40 kDa); and + (nanocarriers at 96 hrs).

Article Snippet: The 4T1 murine breast cancer cell line was a generous gift from Dr. Michael Reiss, The Cancer Institute of New Jersey (New Brunswick, NJ).

Techniques: Ex Vivo, Clinical Proteomics, Comparison