atcc rrid cvcl 0320 17 cl1  (ATCC)

Journal: Journal of Korean Neurosurgical Society

Article Title: Spermine Synthase : A Potential Prognostic Marker for Lower-Grade Gliomas

doi: 10.3340/jkns.2024.0080

Figure Lengend Snippet: Knockdown of spermine synthase (SMS) inhibits the malignant behavior of lower-grade glioma cells. A : Western blot analysis of SMS protein expression levels in human normal glial cell lines CP-H121/122/123 and glioma cell line HS683/U251, with statistical analysis on the right. B : The efficiency of SMS knockdown in HS683/U251 glioma cell line detected by Western blot, with protein expression analysis on the right. C : The efficiency of SMS knockdown in HS683/U251 glioma cell line detected by quantitative reverse transcription polymerase chain reaction. D : Cell proliferation before and after SMS knockdown was measured by CCK-8 assay. E : The apoptosis ratio of cells before and after SMS knockdown was measured by flow cytometry, with statistical analysis on the right. F : Several cell clones formed before and after SMS knockdown, with statistical results. G : Migration of cells before and after SMS knockdown measured by scratch wound healing assay. H : Migration and invasive ability of cells before and after SMS knockdown measured by transwell assay. NC : normal control. *** p <0.001.

Article Snippet: The HS683 (HTB-138) human oligodendroglioma cell line and the U251 (HTB-17) glioblastoma cell line were obtained from ATCC (Manassas, VA, USA).

Techniques: Knockdown, Western Blot, Expressing, Reverse Transcription, Polymerase Chain Reaction, CCK-8 Assay, Flow Cytometry, Clone Assay, Migration, Wound Healing Assay, Transwell Assay, Control

Journal: Scientific reports

Article Title: Assessment of molecular modulation by multifrequency electromagnetic pulses to preferably eradicate tumorigenic cells.

doi: 10.1038/s41598-024-81171-x

Figure Lengend Snippet: Fig. 1. Degree of inhibition of cell viability caused by MEMP as evaluated by an MTT assay. Quantitative determination of cell metabolic activity by MTT in non-treated controls and upon the indicated MEMP treatments. (A) Evaluation of cell viability in U373 MG, HeLa, and MC-38-Luc cancer cell lines treated from 0 to 5 min. (B) Evaluation of viability after MEMP treatment applied for 2.5 and 5 min to HaCaT, CV-1, COS-1, A9 cell lines, and PAM. Data are the mean with standard errors obtained from triplicates and were statistically analyzed by using a one-way ANOVA with a Dunnett’s test (*P < 0.05; **P < 0.01; ****P < 0.0001).

Article Snippet: The origin of our cell lines was as follows: COS-1 (CRL-1650), CV-1 cells (CCL-70) from African green monkey kidney, HEK-293T cells (CRL-3216) (Human embryonic kidney cells), and A9 mouse fibroblasts (CRL-3265), were obtained from the American Type Culture Collection (ATCC); U373 MG (HTB-17) human glioblastoma astrocytoma was purchased from ATCC; the MC-38-Luc cells, derived from MC-38 cell line (SCC172, SigmaAldrich), C57BL/6 murine colon adenocarcinoma cells and stably expressing luciferase gene, were generated in our laboratory as described below; HeLa cells (CCL-2) from human cervical carcinoma were obtained from the ATCC; and the HaCaT human keratinocyte cell line was kindly provided by Dr. Miguel Quintanilla (Instituto de Investigaciones Biomédicas “Alberto Sols”, Madrid, Spain).

Techniques: Inhibition, MTT Assay, Activity Assay

Journal: Scientific reports

Article Title: Assessment of molecular modulation by multifrequency electromagnetic pulses to preferably eradicate tumorigenic cells.

doi: 10.1038/s41598-024-81171-x

Figure Lengend Snippet: Fig. 2. Differential inhibitory effect of MEMP treatments on cell lines assessed by colony forming ability. Control and modulated cell lines at the times indicated in the figure were seeded in triplicate at 103 to 104 cells onto P60 plates. The percentages of colonies formed in culture were calculated with respect to the number of colonies formed in the control (T0). (A) Percentage of colonies formed by U373, HeLa, and MC-38-Luc cells upon the MEMP treatments performed between 2.5 to 5 min. (B) Analysis of the inhibition of colony forming capacity caused by MEMP applied for 2.5 and 5 min to the HaCaT, COS-1, CV-1, and A9 cell lines. (C) Representative photographs showing the effect of MEMP treatment on colony formation by different cell lines. Colonies were fixed and stained with crystal violet two to three weeks after treatments. Data were statistically analyzed by using a one-way ANOVA with Dunnett’s test (*P < 0.05; **P < 0.01; ****P < 0.0001).

Article Snippet: The origin of our cell lines was as follows: COS-1 (CRL-1650), CV-1 cells (CCL-70) from African green monkey kidney, HEK-293T cells (CRL-3216) (Human embryonic kidney cells), and A9 mouse fibroblasts (CRL-3265), were obtained from the American Type Culture Collection (ATCC); U373 MG (HTB-17) human glioblastoma astrocytoma was purchased from ATCC; the MC-38-Luc cells, derived from MC-38 cell line (SCC172, SigmaAldrich), C57BL/6 murine colon adenocarcinoma cells and stably expressing luciferase gene, were generated in our laboratory as described below; HeLa cells (CCL-2) from human cervical carcinoma were obtained from the ATCC; and the HaCaT human keratinocyte cell line was kindly provided by Dr. Miguel Quintanilla (Instituto de Investigaciones Biomédicas “Alberto Sols”, Madrid, Spain).

Techniques: Control, Inhibition, Staining

Journal: Scientific reports

Article Title: Assessment of molecular modulation by multifrequency electromagnetic pulses to preferably eradicate tumorigenic cells.

doi: 10.1038/s41598-024-81171-x

Figure Lengend Snippet: Fig. 3. Determination of cell viability after MEMP treatment by cytometry. Cells modulated at times indicated in the figure were stained with the fixable Viability Dye Ghost Dye Red 780 (1 µg/mL) and fixed. Cells were then analyzed in a FACS Canto flow cytometer (BD Science) to determine the percentage of death and live cells. The figure shows representative data from three experiments on the percentages of live and dead cells after each MEMP modulation condition. Data were statistically analyzed by using a two-way ANOVA with Dunnett’s test (*P < 0.05; **P < 0.01; ****P < 0.0001). (A) Evaluation of MEMP effect on the viability of the U373, HeLa, and MC-38-Luc cancer cells. The experiment was repeated three times. (B) Determination of cell viability by FACS after the MEMP treatment in the HaCaT, CV-1, COS-1, A9 cell lines, and PAM. (C) Indicative photograph of dot plots showing the percentage of death and live U373 and A9 cells.

Article Snippet: The origin of our cell lines was as follows: COS-1 (CRL-1650), CV-1 cells (CCL-70) from African green monkey kidney, HEK-293T cells (CRL-3216) (Human embryonic kidney cells), and A9 mouse fibroblasts (CRL-3265), were obtained from the American Type Culture Collection (ATCC); U373 MG (HTB-17) human glioblastoma astrocytoma was purchased from ATCC; the MC-38-Luc cells, derived from MC-38 cell line (SCC172, SigmaAldrich), C57BL/6 murine colon adenocarcinoma cells and stably expressing luciferase gene, were generated in our laboratory as described below; HeLa cells (CCL-2) from human cervical carcinoma were obtained from the ATCC; and the HaCaT human keratinocyte cell line was kindly provided by Dr. Miguel Quintanilla (Instituto de Investigaciones Biomédicas “Alberto Sols”, Madrid, Spain).

Techniques: Cytometry, Staining, Flow Cytometry

Journal: Scientific reports

Article Title: Assessment of molecular modulation by multifrequency electromagnetic pulses to preferably eradicate tumorigenic cells.

doi: 10.1038/s41598-024-81171-x

Figure Lengend Snippet: Fig. 4. MEMP impact on cell cycle control. (A,B) Cell cycle distribution of malignant U373, HeLa and MC- 38-Luc cell lines, and (C) low tumorigenic A9, HaCaT, CV-1 and COS-1 cell lines, after MEMP modulation treatments ( ). The profiles were obtained by FACS Calibur flow cytometry (BD Science), and cell cycle progression was based on DNA quantification. Cells were modulated for 5 min, then collected at 48 h afterward, fixed and stained with the PI/RNase buffer (BD Pharmingen).

Article Snippet: The origin of our cell lines was as follows: COS-1 (CRL-1650), CV-1 cells (CCL-70) from African green monkey kidney, HEK-293T cells (CRL-3216) (Human embryonic kidney cells), and A9 mouse fibroblasts (CRL-3265), were obtained from the American Type Culture Collection (ATCC); U373 MG (HTB-17) human glioblastoma astrocytoma was purchased from ATCC; the MC-38-Luc cells, derived from MC-38 cell line (SCC172, SigmaAldrich), C57BL/6 murine colon adenocarcinoma cells and stably expressing luciferase gene, were generated in our laboratory as described below; HeLa cells (CCL-2) from human cervical carcinoma were obtained from the ATCC; and the HaCaT human keratinocyte cell line was kindly provided by Dr. Miguel Quintanilla (Instituto de Investigaciones Biomédicas “Alberto Sols”, Madrid, Spain).

Techniques: Control, Flow Cytometry, Staining

Journal: Scientific reports

Article Title: Assessment of molecular modulation by multifrequency electromagnetic pulses to preferably eradicate tumorigenic cells.

doi: 10.1038/s41598-024-81171-x

Figure Lengend Snippet: Fig. 5. Evaluation of MEMP effects on cell shape and actin cytoskeleton organization. Cells were treated with MEMP for two minutes and immediately fixed for subsequent inspection by confocal microscopy with specific antibodies. (A) Fields of A9 and U373 cells stained with the indicated antibody and phalloidin and analyzed by confocal microscopy at 60x magnification. Scale bars, 20 μm. (B) Quantitative determination of the collapse of the actin cytoskeleton as determined by confocal microscopy (see Materials and Methods). Each dot represents a single cell (N, at least180 cells per condition). Statistics was performed using the Brown-Forsythe and Welch ANOVA and the Games-Howell´s multiple comparison tests. (C) Representative picture showing the colony forming ability of A9 and U373 cells at the MEMP regime used in A and B.

Article Snippet: The origin of our cell lines was as follows: COS-1 (CRL-1650), CV-1 cells (CCL-70) from African green monkey kidney, HEK-293T cells (CRL-3216) (Human embryonic kidney cells), and A9 mouse fibroblasts (CRL-3265), were obtained from the American Type Culture Collection (ATCC); U373 MG (HTB-17) human glioblastoma astrocytoma was purchased from ATCC; the MC-38-Luc cells, derived from MC-38 cell line (SCC172, SigmaAldrich), C57BL/6 murine colon adenocarcinoma cells and stably expressing luciferase gene, were generated in our laboratory as described below; HeLa cells (CCL-2) from human cervical carcinoma were obtained from the ATCC; and the HaCaT human keratinocyte cell line was kindly provided by Dr. Miguel Quintanilla (Instituto de Investigaciones Biomédicas “Alberto Sols”, Madrid, Spain).

Techniques: Confocal Microscopy, Staining, Comparison