Associated Anisotropy of intrinsic NAD(P)H for monitoring changes in the metabolic activities of breast cancer cells in three-dimensional collagen matrix

Author(s):  
Anh Cong ◽  
Rafaela M. L. Pimenta ◽  
Jon Holy ◽  
Ahmed A Heikal

The majority of in vitro studies of living cells are routinely conducted in a two-dimensional (2D) monolayer culture. Recent studies, however, suggest that 2D cell culture promotes specific types of...

2016 ◽  
Vol 4 (19) ◽  
pp. 3339-3350 ◽  
Author(s):  
Weijun Xu ◽  
Junmin Qian ◽  
Yaping Zhang ◽  
Aili Suo ◽  
Ning Cui ◽  
...  

Standard two-dimensional monolayer culture platforms often fail to mimic the tumor microenvironment, while animal models complicate research on the effect of individual factors on cell behaviors. Three-dimensional in vitro tumor models bridge gap between them.


PLoS ONE ◽  
2020 ◽  
Vol 15 (12) ◽  
pp. e0243789
Author(s):  
Yonghong Mao ◽  
Yang Zhao ◽  
Yong Zhang ◽  
Hao Yang

Native intact N-glycopeptide analysis can provide access to the comprehensive characteristics of N-glycan occupancy, including N-glycosites, N-glycan compositions, and N-glycoproteins for complex samples. The sample pre-processing method used for the analysis of intact N-glycopeptides usually depends on the enrichment of low abundance N-glycopeptides from a tryptic peptide mixture using hydrophilic substances before LC-MS/MS detection. However, the number of identified intact N-glycopeptides remains inadequate to achieve an in-depth profile of the N-glycosylation landscape. Here, we optimized the sample preparation workflow prior to LC-MS/MS analysis by systematically comparing different analytical methods, including the use of different sources of trypsin, combinations of different proteases, and different enrichment materials. Finally, we found that the combination of Trypsin (B)/Lys-C digestion and zwitterionic HILIC (Zic-HILIC) enrichment significantly improved the mass spectrometric characterization of intact N-glycopeptides, increasing the number of identified intact N-glycopeptides and offering better analytical reproducibility. Furthermore, the optimized workflow was applied to the analysis of intact N-glycopeptides in two-dimensional (2D) and three-dimensional (3D)-cultured breast cancer cells in vitro and xenografted tumors in mice. These results indicated that the same breast cancer cells, when cultured in different microenvironments, can show different N-glycosylation patterns. This study also provides an interesting comparison of the N-glycoproteome of breast cancer cells cultured in different growth conditions, indicating the important role of N-glycosylated proteins in cancer cell growth and the choice of the cell culture model for studies in tumor biology and drug evaluation.


2015 ◽  
Vol 7 (11) ◽  
pp. 1402-1411 ◽  
Author(s):  
Yu Ling Huang ◽  
Chih-kuan Tung ◽  
Anqi Zheng ◽  
Beum Jun Kim ◽  
Mingming Wu

A novel microfluidic model reveals roles of interstitial flows in regulating tumor cell morphology and migration within a 3D collagen matrix. This work highlights the importance of biological flows in tumor cell dissemination.


2013 ◽  
Vol 04 (04) ◽  
pp. 316-326 ◽  
Author(s):  
Guangyao Xiong ◽  
Honglin Luo ◽  
Feng Gu ◽  
Jing Zhang ◽  
Da Hu ◽  
...  

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 ◽  
...  

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).  


2021 ◽  
Vol 2021 ◽  
pp. 1-13
Author(s):  
Xiaowen Wu ◽  
Junxiang Su ◽  
Jizhen Wei ◽  
Nan Jiang ◽  
Xuejun Ge

Cell culture is one of the most core and fundamental techniques employed in the fields of biology and medicine. At present, although the two-dimensional cell culture method is commonly used in vitro, it is quite different from the cell growth microenvironment in vivo. In recent years, the limitations of two-dimensional culture and the advantages of three-dimensional culture have increasingly attracted more and more attentions. Compared to two-dimensional culture, three-dimensional culture system is better to realistically simulate the local microenvironment of cells, promote the exchange of information among cells and the extracellular matrix (ECM), and retain the original biological characteristics of stem cells. In this review, we first present three-dimensional cell culture methods from two aspects: a scaffold-free culture system and a scaffold-based culture system. The culture method and cell characterizations will be summarized. Then the application of three-dimensional cell culture system is further explored, such as in the fields of drug screening, organoids and assembloids. Finally, the directions for future research of three-dimensional cell culture are stated briefly.


2021 ◽  
Vol 2021 ◽  
pp. 1-14
Author(s):  
F. Beheshti ◽  
A. A. Shabani ◽  
M. R. Akbari Eidgahi ◽  
P. Kookhaei ◽  
M. Vazirian ◽  
...  

Cancer is a leading cause of death and a vital health care challenge in the world. Hence, this work was conducted to determine the in vitro anticancer property and also the molecular mechanism of aqueous and organic extracts of Ipomoea purpurea leaves in three human cancer cell lines, including A-549 (human lung cancer), HepG-2 (human liver cancer), MDA-MB-231 (human breast cancer), and MCF-10A (breast normal cell line). In vitro cytotoxic potential of organic extracts, such as hexane, chloroform, ethyl-acetate, methanol, and aqueous extract was examined using a standard (3-(4,5-dimethylthiazole)-2,5-diphenyltetrazolium bromide) MTT method in both monolayer two-dimensional (2D) and spheroids multicellular three-dimensional (3D) cultures. The MTT assay data showed that methanol and chloroform extracts of I. purpurea leaves had the antiproliferative effect on lung and breast cancer cells with IC50 of 53.62 ± 0.07 and 124.5 ± 0.01 µg/mL, respectively. The results of further examinations, such as dual acridine orange/ethidium bromide, Annexin V-FITC/PI, and caspase-3 colorimetric assay, confirmed that methanol and chloroform extracts of I. purpurea as the most potent cytotoxic extracts might contain a variety of phytochemicals, promoting apoptosis in lung and breast cancer cells. The present research findings suggested that methanolic extract of I. purpurea leaves induced S-phase cell cycle arrest and intrinsic pathway of apoptosis in A-549 lung cancer cells. The study further showed that I. purpurea could be a helpful candidate for cancer treatment.


2021 ◽  
Author(s):  
Sangeeta Ballav ◽  
Ankita Jaywant Deshmukh ◽  
Shafina Siddiqui ◽  
Jyotirmoi Aich ◽  
Soumya Basu

Cell culture is one of the most important and commonly used in vitro tools to comprehend various aspects of cells or tissues of a living body such as cell biology, tissue morphology, mechanism of diseases, cell signaling, drug action, cancer research and also finds its great importance in preclinical trials of various drugs. There are two major types of cell cultures that are most commonly used- two-dimensional (2D) and three-dimensional culture (3D). The former has been used since the 1900s, owing to its simplicity and low-cost maintenance as it forms a monolayer, while the latter being the advanced version and currently most worked upon. This chapter intends to provide the true meaning and significance to both cultures. It starts by making a clear distinction between the two and proceeds further to discuss their different applications in vitro. The significance of 2D culture is projected through different assays and therapeutic treatment to understand cell motility and treatment of diseases, whereas 3D culture includes different models and spheroid structures consisting of multiple layers of cells, and puts a light on its use in drug discovery and development. The chapter is concluded with a detailed account of the production of therapeutic proteins by the use of cells.


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