Targeted metabolomics in the cell culture media reveals increased uptake of branched amino acids by breast cancer cells

2021 ◽  
pp. 114192
Author(s):  
Fang Kou ◽  
Bangjie Zhu ◽  
Wenbin Zhou ◽  
Chunming Lv ◽  
Yu Cheng ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Amino Acids ◽  
Cell Culture ◽  
Cancer Cells ◽  
Breast Cancer Cells ◽  
Culture Media ◽  
Targeted Metabolomics ◽  
Cell Culture Media ◽  
Branched Amino Acids

Targeted metabolomics reveals dynamic portrayal of amino acids and derivatives in triple-negative breast cancer cells and culture media

2021 ◽  
Author(s):  
Fang Kou ◽  
Bangjie Zhu ◽  
Wenbin Zhou ◽  
Chunming Lv ◽  
Yu Cheng ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Amino Acids ◽  
Cancer Cells ◽  
Triple Negative Breast Cancer ◽  
Breast Cancer Cells ◽  
Triple Negative ◽  
Culture Media ◽  
Targeted Metabolomics ◽  
Amino Acids And Derivatives

Targeted metabolomics reveals dynamic portrayal of amino acids and derivatives in triple-negative breast cancer cells and culture media.

scholarly journals Sulforaphane Modulates Cytokine Levels in Cell Culture Media of Triple-Negative Breast Cancer Cells Grown with Macrophages

2020 ◽  
Vol 4 (Supplement_2) ◽  
pp. 337-337
Author(s):  
Courtney Merrick ◽  
Lauren Housley
Keyword(s):  
Breast Cancer ◽  
Cell Culture ◽  
Breast Cancer Cells ◽  
Triple Negative ◽  
Culture Media ◽  
California State University ◽  
State University ◽  
Cell Culture Media ◽  
Cytokine Levels ◽  
The Media

Abstract Objectives Triple negative breast cancer (TNBC) makes up approximately 10–20% of all breast cancer cases and is more common in younger women and in Hispanic and African American populations. It is particularly difficult to treat, exhibiting high-metastasis rates, poor prognosis, and limited treatment options. Mortality from TNBC is largely due to the tumor cells high invasive capacity and rapid progression to metastasis. Evidence suggests that macrophages in the breast tumor microenvironment release cytokines that increase tumor cell proliferation, invasion and metastasis. Sulforaphane (SFN) is a broccoli phytochemical that has been identified to slow the progression of breast cancer as well as alter cytokine secretion from macrophages and breast cancer cells grown in single culture. SFN effects on cytokine secretion in the breast tumor microenvironment remain unclear. This study is investigating the effect of SFN on cytokine levels in cell culture media of TNBC cells grown with and without macrophages. Our hypothesis is that cytokine levels differ in media from cocultured cells versus singly cultured cells, and SFN treatment further alters cytokine levels in media. Methods In this study, TNBC cells (MDA-MB-231) were grown in transwell plates with and without macrophages (THP-1 cells differentiated with phorbol-myristate acetate). Cell cultures (n = 3) were treated with either 15 µM SFN, DMSO (vehicle-control), or a non-treatment control. We evaluated the levels of 44 individual cytokines in cell culture media at 24 and 48 hours after treatment using a multi-plex (BioPlex) assay. Control groups included single-cultured MDA-MB-231 and differentiated THP-1 cells. Results Preliminary analyses revealed that cytokine levels differed in the media of single versus cocultured cells and among treatment groups after 24 and 48 hours of treatment. Conclusions The profile of cytokines in the media of TNBC cells grown with macrophages was influenced by SFN treatment. This information may help establish mechanisms underlying SFN effects on TNBC behavior and identify new treatment strategies. Funding Sources California State University Program for Education and Research in Biotechnology, California State University-Chico.

scholarly journals Effect of Modification Protocols on the Effectiveness of Gold Nanoparticles as Drug Delivery Vehicles for Killing of Breast Cancer Cells

2016 ◽  
Vol 69 (12) ◽  
pp. 1402 ◽  
Author(s):  
Zahrah Alhalili ◽  
Daniela Figueroa ◽  
Martin R. Johnston ◽  
Joe Shapter ◽  
Barbara Sanderson
Keyword(s):  
Breast Cancer ◽  
Cell Culture ◽  
Gold Nanoparticles ◽  
Cancer Cells ◽  
Breast Cancer Cells ◽  
Culture Media ◽  
High Dose ◽  
Dependent Manner ◽  
T47d Cells

The current study evaluated the potential of gold nanoparticles (AuNPs) for the delivery of Taxol to breast cancer cells (T47D) using an in vitro cell culture model. For this study, new loading approaches and novel chemical attachments were investigated. Five different gold nanoparticle-based complexes were used to determine their cytotoxicity towards T47D cells using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) viability assay. There was no significant decrease (P > 0.05) in cell viability when T47D cells were treated with AuNPs that did not contain Taxol. However, cells were significantly killed by gold nanoparticles chemically conjugated to Taxol using three different approaches and one novel hybrid AuNP-Taxol nanoparticle, wherein no chemical bonds were involved. These Taxol-loaded AuNPs were more effective at inducing cell death in vitro than a solution of free Taxol used to treat cells. This result demonstrated that Taxol could be released from the particles in the cell culture media for subsequent therapeutic action. Additionally, the experiments proved that the Taxol-loaded AuNPs were more toxic in a dose dependent manner than Taxol as a formulation for the treatment of breast cancer cells. The results of this study suggest that gold nanoparticles have potential for the efficient delivery of Taxol to breast cancer cells. This could provide a future solution as an alternative application method to overcome adverse side effects resulting from current high-dose treatment regimes.

Length vs. stiffness: which plays a dominant role in the cellular uptake of fructose-based rod-like micelles by breast cancer cells in 2D and 3D cell culture models?

2018 ◽  
Vol 6 (25) ◽  
pp. 4223-4231 ◽  
Author(s):  
Jiacheng Zhao ◽  
Hongxu Lu ◽  
Yin Yao ◽  
Sylvia Ganda ◽  
Martina H. Stenzel
Keyword(s):  
Breast Cancer ◽  
Cell Culture ◽  
Cancer Cells ◽  
Breast Cancer Cells ◽  
Dominant Role ◽  
3D Cell Culture ◽  
Culture Models ◽  
Nano Rods ◽  
2D And 3D ◽  
Cell Culture Models

Internalization of rod-like micelles by breast cancer cells is significantly affected by the stiffness of nano-rods.

Effects of the Gold Nanoparticles (AuNPs) on the Proliferation and Morphological Characteristics of Human Breast Cancer Cells (MCF-7) in Culture

2017 ◽  
Vol 268 ◽  
pp. 254-258 ◽  
Author(s):  
Nur Shafawati Rosli ◽  
Azhar Abdul Rahman ◽  
Azlan Abdul Aziz ◽  
Shaharum Shamsuddin ◽  
Nurul Sabihah Zakaria
Keyword(s):  
Breast Cancer ◽  
Cancer Cells ◽  
Breast Cancer Cells ◽  
Culture Media ◽  
Morphological Changes ◽  
Morphological Characteristics ◽  
Morphological Observation ◽  
Duration Of Treatment ◽  
Significant Difference ◽  
Mcf 7

Ultrastructural characteristic and morphological changes of untreated and treated breast cancer MCF-7 cells were observed by energy-filtered transmission electron microscope (EFTEM). Morphological observation of MCF-7 after being treated with 13 nm, 50 nm, and 70 nm AuNPs, were looking unhealthy and dying out of the populace, the observed cells were more reduced and dying as treatment with 50 nm and 70 nm AuNPs. Cells detachment, clumping, shrunken, and dispersed cells in the culture medium and floating cells were also observed. The observed morphological changes increase in 50 nm and 70 nm AuNPs than in 13 nm AuNPs, which is less toxic to MCF-7 cells. The presented morphological analysis has established that 13 nm AuNPs showed less toxic to MCF-7 breast cancer cells. Whereas, control cells of MCF-7 were treated with only complete culture media, despite the duration of treatment, whereby the cells maintained most of their morphological features and observed to have a typical morphology of healthy cells that are well attached to the surface. These results indicate that AuNPs were clustered in the cells and there was no significant difference between images of different sizes of AuNPs observed in the cells, because the AuNPs always clustered together inside the cells.

scholarly journals A big cheese in biotherapeutics: Lactoyl leucine and isoleucine are bioavailable alternatives for canonical amino acids in cell culture media

2021 ◽  
Author(s):  
Corinna Schmidt ◽  
Maria Wehsling ◽  
Maxime Le Mignon ◽  
Gregor Wille ◽  
Yannick Rey ◽  
...  
Keyword(s):  
Amino Acids ◽  
Cell Culture ◽  
Cho Cells ◽  
Culture Media ◽  
Chinese Hamster ◽  
Batch Processes ◽  
Next Generation ◽  
Cell Culture Media ◽  
Derivatives Of

Increasing demands for protein-based therapeutics such as monoclonal antibodies, fusion proteins, bispecific molecules and antibody fragments require researchers to constantly find innovative solutions. To increase yields and decrease costs of next generation bioprocesses, highly concentrated cell culture media formulations are developed but often limited by the low solubility of amino acids such as tyrosine, cystine, leucine and isoleucine, in particular at physiological pH. This work sought to investigate highly soluble and bioavailable derivatives of leucine and isoleucine that are applicable for fed-batch processes. N-lactoyl-leucine and N-lactoyl-isoleucine sodium salts were tested in cell culture media and proved to be beneficial to increase the overall solubility of cell culture media formulations. These modified amino acids proved to be bioavailable for various Chinese hamster ovary (CHO) cells and were suitable for replacement of canonical amino acids in cell culture feeds. The quality of the final recombinant protein was studied in bioprocesses using the derivatives, and the mechanism of cleavage was investigated in CHO cells. Altogether, both N-lactoyl amino acids represent an advantageous alternative to canonical amino acids to develop highly concentrated cell culture media formulations to support next generation bioprocesses.

scholarly journals Anti-tumor activity of plant extracts against human breast cancer cells are different in monolayer and three-dimensional cell culture screening models: A comparison on 34 extracts

2020 ◽  
Vol 7 (3) ◽  
pp. 3667-3677
Author(s):  
Nhan Lu-Chinh Phan ◽  
Khuong Duy Pham ◽  
Mai Thi-Thanh Nguyen ◽  
Ngoc Kim Phan ◽  
Kiet Dinh Truong ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Cell Culture ◽  
Cancer Cells ◽  
Cell Lines ◽  
Plant Extracts ◽  
Breast Cancer Cells ◽  
Three Dimensional ◽  
Culture Model ◽  
Anti Tumor Activity ◽  
Mcf 7

Introduction: The monolayer cell culture model is a popular model for screening anti-tumor activity of plant extracts. However, almost the extracts selected for screening in this model have failed in subsequent animal models. Therefore, there is only about 5 % of candidates from the original thousands of drugs that are screened which ultimately reach clinical trial. This study aimed to compare the differences in anti-tumor activity of 34 plant extracts against breast cancer cells in 2 models of monolayer cell culture (2D) and in three-dimensional (3D) cell culture. Methods: Four breast cancer cell lines (MCF-7, CD44+CD24- MCF-7, VN9, and CD44+CD24- VN9) were used to generate the 2D and 3D models (the 3D model was developed by culturing breast cancer cells in matrigel). The extracts were got from the plant extract library that prepared in the previous study. The anti-tumor activity was evaluated via half inhibitory concentrations( IC50 values). Results: Of the 34 extracts, E12, E7, E5 and E6 of them had an effect on MCF-7, CD44+CD24- MCF-7, VN9 and CD44+CD24- VN9 cells, respectively. The results indicated 10 potentially strong candidates for future drug development targeting hypoxic areas in breast cancer. Conclusion: The 3D culture model exhibited higher resistance to extracts than the 2D culture model. The CD44+CD24- cell population of both VN9 and MCF-7 cell lines showed higher drug resistance than the original cell lines (VN9 and MCF-7).  

scholarly journals Typhonium flagelliforme decreases telomerase expression in HeLa cervical cancer cells

2016 ◽  
Vol 35 (1) ◽  
pp. 3
Author(s):  
Endang Purwaningsih ◽  
Yulia Suciati ◽  
Etty Widayanti
Keyword(s):  
Breast Cancer ◽  
Cervical Cancer ◽  
Cancer Cells ◽  
Breast Cancer Cells ◽  
Culture Media ◽  
Vero Cells ◽  
Telomerase Activity ◽  
Multiple Comparison Test ◽  
Cervical Cancer Cells ◽  
Experimental Laboratory

<p><strong>Background</strong><br /> Cancer cells have a relatively high telomerase activity compared to normal cells, so that cancer cells have the ability for continued proliferation and uncontrolled mitosis. Telomerase is an enzyme responsible for the length of telomeres, DNA segments located at the ends of eukaryotic chromosomes. Natural materials such as rodent tuber (<em>Typhonium flagelliforme</em>) have anticancer potential. The purpose of the present study was to determine the effects of <em>Typhonium flagelliforme </em>extract on telomerase expression in HeLa cervical cancer and T47D breast cancer cells.</p><p><br /> <strong>Methods </strong></p><p>This experimental laboratory study was conducted on cultured HeLa and T47D cancer cell lines, with normal Vero cells as controls, and using RPMI and M199 culture media. The study comprised three groups, i.e. controls, and groups receiving <em>Typhonium flagelliforme </em>extract at doses of ½ IC50 and IC50. Telomerase expression was measured by immunohistochemistry (IHC). Analysis of variance and LSD multiple comparison test were used to analyze the data.</p><p><strong> </strong></p><p><strong>Results </strong><br /> Telomerase expression in cancer cells showed significantly higher values compared to normal Vero cells. <em>Typhonium flagelliforme</em> extract was capable of significantly decreasing telomerase expression in cancer cells receiving the extract.</p><p><strong> </strong></p><strong>Conclusion </strong><br /> <em>Typhonium flagelliforme</em> extract at different doses is capable of decreasing telomerase expression more effectively in cervical cancer cells than in breast cancer cells. This study shows that <em>Typhonium flagelliforme</em> may have anti-cancer activity, necessitating further investigations.

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