scholarly journals Screening of Additive Manufactured Scaffolds Designs for Triple Negative Breast Cancer 3D Cell Culture and Stem-Like Expansion

2018 ◽  
Vol 19 (10) ◽  
pp. 3148 ◽  
Author(s):  
Emma Polonio-Alcalá ◽  
Marc Rabionet ◽  
Antonio Guerra ◽  
Marc Yeste ◽  
Joaquim Ciurana ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Cell Culture ◽  
Porous Scaffold ◽  
Three Dimensional ◽  
3D Cell Culture ◽  
Poly Lactic Acid ◽  
Tumor Initiating Cells ◽  
Fused Filament Fabrication ◽  
Open Issue

Breast cancer stem cells (BCSCs) are tumor-initiating cells responsible for metastasis and tumor reappearance, but their research is limited by the impossibility to cultivate them in a monolayer culture. Scaffolds are three-dimensional (3D) cell culture systems which avoid problems related with culturing BCSC. However, a standardized scaffold for enhancing a BCSC population is still an open issue. The main aim of this study is to establish a suitable poly (lactic acid) (PLA) scaffold which will produce BCSC enrichment, thus allowing them to be studied. Different 3D printing parameters were analyzed using Taguchi experimental design methods. Several PLA scaffold architectures were manufactured using a Fused Filament Fabrication (FFF) 3D printer. They were then evaluated by cell proliferation assay and the configurations with the highest growth rates were subjected to BCSC quantification by ALDH activity. The design SS1 (0.2 mm layer height, 70% infill density, Zigzag infill pattern, 45° infill direction, and 100% flow) obtained the highest proliferation rate and was capable of enhancing a ALDH+ cell population compared to 2D cell culture. In conclusion, the data obtained endorse the PLA porous scaffold as useful for culturing breast cancer cells in a microenvironment similar to in vivo and increasing the numbers of BCSCs.

scholarly journals Mammary gland 3D cell culture systems in farm animals

2021 ◽  
Vol 52 (1) ◽  
Author(s):  
Laurence Finot ◽  
Eric Chanat ◽  
Frederic Dessauge
Keyword(s):  
Cell Culture ◽  
Mammary Gland ◽  
Bacterial Infections ◽  
Three Dimensional ◽  
3D Cell Culture ◽  
Farm Animals ◽  
Culture Systems ◽  
Cell Culture Systems

AbstractIn vivo study of tissue or organ biology in mammals is very complex and progress is slowed by poor accessibility of samples and ethical concerns. Fortunately, however, advances in stem cell identification and culture have made it possible to derive in vitro 3D “tissues” called organoids, these three-dimensional structures partly or fully mimicking the in vivo functioning of organs. The mammary gland produces milk, the source of nutrition for newborn mammals. Milk is synthesized and secreted by the differentiated polarized mammary epithelial cells of the gland. Reconstructing in vitro a mammary-like structure mimicking the functional tissue represents a major challenge in mammary gland biology, especially for farm animals for which specific agronomic questions arise. This would greatly facilitate the study of mammary gland development, milk secretion processes and pathological effects of viral or bacterial infections at the cellular level, all with the objective of improving milk production at the animal level. With this aim, various 3D cell culture models have been developed such as mammospheres and, more recently, efforts to develop organoids in vitro have been considerable. Researchers are now starting to draw inspiration from other fields, such as bioengineering, to generate organoids that would be more physiologically relevant. In this chapter, we will discuss 3D cell culture systems as organoids and their relevance for agronomic research.

scholarly journals A reliable and affordable 3D tumor spheroid model for natural product drug discovery: A case study of curcumin

2019 ◽  
Vol 2 (1) ◽  
Author(s):  
Loh Teng Hern Tan ◽  
Liang Ee Low ◽  
Siah Ying Tang ◽  
Wei Hsum Yap ◽  
Lay Hong Chuah ◽  
...  
Keyword(s):  
Cell Culture ◽  
Drug Discovery ◽  
Three Dimensional ◽  
3D Cell Culture ◽  
Automated Image Analysis ◽  
Tumor Spheroids ◽  
Culture Methods ◽  
In Vivo Models

Three-dimensional cell culture methods revolutionize the field of anticancer drug discovery, forming an important link-bridge between conventional in vitro and in vivo models and conferring significant clinical and biological relevant data. The current work presents an affordable yet reproducible method of generating homogenous 3D tumor spheroids. Also, a new open source software is adapted to perform an automated image analysis of 3D tumor spheroids and subsequently generate a list of morphological parameters of which could be utilized to determine the response of these spheroids toward treatments. Our data showed that this work could serve as a reliable 3D cell culture platform for preclinical cytotoxicity testing of natural products prior to the expensive and time-consuming animal models

Application of Three-dimensional (3D) Tumor Cell Culture Systems and Mechanism of Drug Resistance

2019 ◽  
Vol 25 (34) ◽  
pp. 3599-3607 ◽  
Author(s):  
Adeeb Shehzad ◽  
Vijaya Ravinayagam ◽  
Hamad AlRumaih ◽  
Meneerah Aljafary ◽  
Dana Almohazey ◽  
...  
Keyword(s):  
Cell Culture ◽  
Cancer Cells ◽  
Anticancer Drugs ◽  
Three Dimensional ◽  
3D Culture ◽  
3D Cell Culture ◽  
Cancer Therapeutics ◽  
In Vivo Model

: The in-vitro experimental model for the development of cancer therapeutics has always been challenging. Recently, the scientific revolution has improved cell culturing techniques by applying three dimensional (3D) culture system, which provides a similar physiologically relevant in-vivo model for studying various diseases including cancer. In particular, cancer cells exhibiting in-vivo behavior in a model of 3D cell culture is a more accurate cell culture model to test the effectiveness of anticancer drugs or characterization of cancer cells in comparison with two dimensional (2D) monolayer. This study underpins various factors that cause resistance to anticancer drugs in forms of spheroids in 3D in-vitro cell culture and also outlines key challenges and possible solutions for the future development of these systems.

scholarly journals 3D Cell Culture Models in COVID-19 Times: A Review of 3D Technologies to Understand and Accelerate Therapeutic Drug Discovery

Biomedicines ◽  
2021 ◽  
Vol 9 (6) ◽  
pp. 602
Author(s):  
Guadalupe Tonantzin de Dios-Figueroa ◽  
Janette del Rocío Aguilera-Marquez ◽  
Tanya A. Camacho-Villegas ◽  
Pavel H. Lugo-Fabres
Keyword(s):  
Cell Culture ◽  
Drug Evaluation ◽  
Three Dimensional ◽  
3D Cell Culture ◽  
Therapeutic Drug ◽  
Emerging Viruses ◽  
Culture Models ◽  
Tissue Characteristics ◽  
Cell Culture Models

In the last decades, emerging viruses have become a worldwide concern. The fast and extensive spread of the disease caused by SARS-CoV-2 (COVID-19) has impacted the economy and human activity worldwide, highlighting the human vulnerability to infectious diseases and the need to develop and optimize technologies to tackle them. The three-dimensional (3D) cell culture models emulate major tissue characteristics such as the in vivo virus–host interactions. These systems may help to generate a quick response to confront new viruses, establish a reliable evaluation of the pathophysiology, and contribute to therapeutic drug evaluation in pandemic situations such as the one that humanity is living through today. This review describes different types of 3D cell culture models, such as spheroids, scaffolds, organoids, and organs-on-a-chip, that are used in virus research, including those used to understand the new severe acute respiratory syndrome-coronavirus 2 (SARS-CoV-2).

scholarly journals THU0058 ENHANCEMENT OF CARTILAGE REGENERATION EFFICIENCY WITH HUMAN ADIPOSE STEM CELL THREE-DIMENSIONAL SPHEROID

2020 ◽  
Vol 79 (Suppl 1) ◽  
pp. 241.2-241
Author(s):  
J. Y. Ko ◽  
E. Lee ◽  
J. Kim ◽  
G. I. Im
Keyword(s):  
Stem Cells ◽  
Cell Culture ◽  
Single Cells ◽  
Three Dimensional ◽  
3D Cell Culture ◽  
Cartilage Regeneration ◽  
Blue Staining ◽  
Alcian Blue Staining ◽  
Culture Dish

Background:3D (three-dimensional) cell culture technology has been researched steadily because of its high potential of biocompatibility compared to single cells since 1990s, and is being developed to 3D spheroids recently. Spheroids are considered to reflect the natural organization of cells better than 2D cell cultures, and stem cells spheroids have been studied extensively in therapeutic transplantation. Stem cells were considered as a method of replacing autologous chondrocyte in regenerative treatment of articular cartilage. Compared to conventional single cells, 3D cell culture is artificially created an environment similar to a living body in vitro so that all cells collectively, a cell culture model that allows growth or interaction with the environment. Therefore, the findings of this study indicate that enhancement of treatment efficiency of stem cells caused by potential of survival and proliferation of hASC spheroid in Osteoarthritis. In conclusion, spheroid positive subpopulation of hASCs has high cell proliferation and survival but not apoptosis and cell death potential, which may contribute to successful cartilage regeneration and the development of stem cell therapies in the future.Objectives:Studied for 3D spheroids to investigate the mechanism of enhancement of survival and proliferationof hASC (human adipose stem cells) spheroid, which may contribute to successful improvement of therapeutic efficacy of stem cells.Methods:Cell isolation and culture / 3D cell culture dish preparation / hASCs culture on 3D cell culture dish / Real-time PCR analysis / Western blotting / Alcian blue staining / ACLT + MM (Anterior cruciate ligament transection with Medial meniscectomy) model / In vivo fluorescence for cell tracking / In vivo effects of spheroids in OA joint / Histological analysis / Enzyme-linked immunosorbent assay (ELISA) results for inflamma -tory cytokines in rat synovial fluid / Statistical AnalysisResults:In order to see how the spheroid showed more residual than single, and how effective it was in actual cartilage regeneration, the result of paraffin tissues were confirmed by safranin O staining for each condition. The tendency of cartilage regeneration efficiency was good for spheroid. Although the differences between the single and spheroid groups were small, they reaffirmed that they could somewhat protect cartilage and help regeneration treatment. However, immunohistochemistry of HN(Human nucleic antigen) staining showed that cells of single and spheroid were not observed in the wound but disappeared by the paracrine effect.Conclusion:Spheroids do not exhibit differentiation characteristics, but they could be seen as a result of expression of related genes such as Bax, Bcl-XL and Alcian blue staining. Spheroids tend to have low potential of cell death rather than proliferation and reduction in the proliferation. So, we conclude the fact that instead of hASCs going directly to the surgical site to regenerate cartilage, they can help catrilage regeneration.Acknowledgments:This research was supported by the National Research Foundation of Korea (NRF-2019R1H1A2039685 and 2019R1I1A1A01043778).Disclosure of Interests:None declared

scholarly journals Activity of trastuzumab emtansine (T-DM1) in 3D cell culture

2021 ◽  
Author(s):  
Jean Zheng Boyer ◽  
Gail D. Lewis Phillips ◽  
Hiro Nitta ◽  
Karl Garsha ◽  
Brittany Admire ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Cell Culture ◽  
Cell Lines ◽  
3D Cell Culture ◽  
Breast Cancer Cell Lines ◽  
Trastuzumab Emtansine ◽  
Culture Methods ◽  
Drug Activity ◽  
Gene And Protein Expression

Abstract Background Cell spheroids and aggregates generated from three-dimensional (3D) cell culture methods are similar to in vivo tumors in terms of tissue morphology, biology, and gene expression, unlike cells grown in 2D cell cultures. Breast cancer heterogeneity is one of the main drug resistant mechanisms and needs to be overcome in order to increase the efficacy of drug activity in its treatments. Methods We performed a unique 3D cell culture and drug efficacy study with trastuzumab emtansine (Kadcyla®, T-DM1) across five breast cancer cell lines (BT-474, SK-BR-3, MDA-MB-361, MDA-MB-175, and MCF-7) that were previously investigated in 2D cell culture. We performed HER2 IHC staining, cell viability experiments, Gene-protein-assay (GPA), and T-DM1 internalization studies. Results We obtained significantly different results including higher IC50 for some of the cell lines. Our GPA showed some significant heterogeneous HER2 gene and protein expression in 3D cultured spheroids or aggregates. The fluorescent images also showed that a longer incubation time is needed for T-DM1 to be internalized effectively into 3D cultured spheroids or aggregates. Conclusion Our study demonstrated that the difference of T-DM1 drug activity in 3D spheroids or aggregates might be due to tumor heterogeneity and less efficient internalization of T-DM1 that is not seen using 2D cell culture models. Drug studies using 3D cell culture are expected to provide biologically relevant models for determining drug activity in tumor tissue in future drug response and resistance research.

scholarly journals PLA Electrospun Scaffolds for Three-Dimensional Triple-Negative Breast Cancer Cell Culture

Polymers ◽  
2019 ◽  
Vol 11 (5) ◽  
pp. 916 ◽  
Author(s):  
Emma Polonio-Alcalá ◽  
Marc Rabionet ◽  
Xavier Gallardo ◽  
David Angelats ◽  
Joaquim Ciurana ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Gene Expression ◽  
Cell Culture ◽  
Triple Negative Breast Cancer ◽  
Triple Negative ◽  
Breast Cancer Subtype ◽  
Three Dimensional ◽  
Poly Lactic Acid ◽  
Mesenchymal Transition ◽  
3D Cultures

Three-dimensional (3D) systems provide a suitable environment for cells cultured in vitro since they reproduce the physiological conditions that traditional cell culture supports lack. Electrospinning is a cost-effective technology useful to manufacture scaffolds with nanofibers that resemble the extracellular matrix that surround cells in the organism. Poly(lactic acid) (PLA) is a synthetic polymer suitable for biomedical applications. The main objective of this study is to evaluate electrospun (ES)-PLA scaffolds to be used for culturing cancer cells. Triple-negative breast cancer (TNBC) is the most aggressive breast cancer subtype with no validated targeted therapy and a high relapse rate. MDA-MB-231 TNBC cells were grown in scaffolds from two different PLA concentrations (12% and 15% w/v). The appropriateness of ES-PLA scaffolds was evaluated using a cell proliferation assay. EGFR and STAT3 gene expression and protein levels were compared in cells grown in 2D versus in 3D cultures. An increase in STAT3 activation was shown, which is related to self-renewal of cancer stem cells (CSCs). Therefore, the enrichment of the breast CSC (BCSC) population was tested using a mammosphere-forming assay and gene expression of BCSC-related stemness and epithelial-to-mesenchymal transition markers. Based on the results obtained, ES-PLA scaffolds are useful for 3D cultures in short culture periods with no BCSC-enrichment.

Natural Polymers Based Hydrogels for Cell Culture Applications

2020 ◽  
Vol 27 (16) ◽  
pp. 2734-2776 ◽  
Author(s):  
Gils Jose ◽  
K.T. Shalumon ◽  
Jyh-Ping Chen
Keyword(s):  
Cell Culture ◽  
Three Dimensional ◽  
3D Cell Culture ◽  
Biological Properties ◽  
Special Focus ◽  
Natural Polymers ◽  
Cell Functions ◽  
Defined Culture

It is well known that the extracellular matrix (ECM) plays a vital role in the growth, survival and differentiation of cells. Though two-dimensional (2D) materials are generally used as substrates for the standard in vitro experiments, their mechanical, structural, and compositional characteristics can alter cell functions drastically. Many scientists reported that cells behave more natively when cultured in three-dimensional (3D) environments than on 2D substrates, due to the more in vivo-like 3D cell culture environment that can better mimic the biochemical and mechanical properties of the ECM. In this regard, water-swollen network polymer-based materials called hydrogels are highly attractive for developing 3D ECM analogs due to their biocompatibility and hydrophilicity. Since hydrogels can be tuned and altered systematically, these materials can function actively in a defined culture medium to support long-term self-renewal of various cells. The physico-chemical and biological properties of the materials used for developing hydrogel should be tunable in accordance with culture needs. Various types of hydrogels derived either from natural or synthetic origins are currently being used for cell culture applications. In this review, we present an overview of various hydrogels based on natural polymers that can be used for cell culture, irrespective of types of applications. We also explain how each hydrogel is made, its source, pros and cons in biological applications with a special focus on regenerative engineering.

scholarly journals Biopolymer-Based Microcarriers for Three-Dimensional Cell Culture and Engineered Tissue Formation

2020 ◽  
Vol 21 (5) ◽  
pp. 1895 ◽  
Author(s):  
Lixia Huang ◽  
Ahmed M.E. Abdalla ◽  
Lin Xiao ◽  
Guang Yang
Keyword(s):  
Cell Culture ◽  
Three Dimensional ◽  
3D Cell Culture ◽  
Cellular Interactions ◽  
Regeneration Medicine ◽  
Promising Tool ◽  
Engineered Tissues ◽  
Engineered Tissue ◽  
And Function

The concept of three-dimensional (3D) cell culture has been proposed to maintain cellular morphology and function as in vivo. Among different approaches for 3D cell culture, microcarrier technology provides a promising tool for cell adhesion, proliferation, and cellular interactions in 3D space mimicking the in vivo microenvironment. In particular, microcarriers based on biopolymers have been widely investigated because of their superior biocompatibility and biodegradability. Moreover, through bottom-up assembly, microcarriers have opened a bright door for fabricating engineered tissues, which is one of the cutting-edge topics in tissue engineering and regeneration medicine. This review takes an in-depth look into the recent advancements of microcarriers based on biopolymers—especially polysaccharides such as chitosan, chitin, cellulose, hyaluronic acid, alginate, and laminarin—for 3D cell culture and the fabrication of engineered tissues based on them. The current limitations and potential strategies were also discussed to shed some light on future directions.

scholarly journals Advances in 3D peptide hydrogel models in cancer research

2021 ◽  
Vol 5 (1) ◽  
Author(s):  
Jingwen Xu ◽  
Guangyan Qi ◽  
Weiqun Wang ◽  
Xiuzhi Susan Sun
Keyword(s):  
Cell Culture ◽  
Food Safety ◽  
Three Dimensional ◽  
Safety Evaluation ◽  
3D Cell Culture ◽  
3D Models ◽  
3D Microenvironment ◽  
Peptide Hydrogel

AbstractIn vitro cell culture models on monolayer surfaces (2D) have been widely adapted for identification of chemopreventive food compounds and food safety evaluation. However, the low correlation between 2D models and in vivo animal models has always been a concern; this gap is mainly caused by the lack of a three-dimensional (3D) extracellular microenvironment. In 2D models, cell behaviors and functionalities are altered, resulting in varied responses to external conditions (i.e., antioxidants) and hence leading to low predictability. Peptide hydrogel 3D scaffolding technologies, such as PGmatrix for cell culture, have been recently reported to grow organoid-like spheroids physiologically mimicking the 3D microenvironment that can be used as an in vitro 3D model for investigating cell activities, which is anticipated to improve the prediction rate. Thus, this review focuses on advances in 3D peptide hydrogels aiming to introduce 3D cell culture tools as in vitro 3D models for cancer-related research regarding food safety and nutraceuticals.

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