scholarly journals Critical role of three-dimensional tumorsphere size on experimental outcome

BioTechniques ◽  
2020 ◽  
Vol 69 (5) ◽  
pp. 333-338
Author(s):  
Suraj Kumar Singh ◽  
Sabiya Abbas ◽  
Ajit Kumar Saxena ◽  
Swasti Tiwari ◽  
Lokendra Kumar Sharma ◽  
...  
Keyword(s):  
Drug Treatment ◽  
Cell Density ◽  
Drug Toxicity ◽  
Critical Role ◽  
Tumor Biology ◽  
Three Dimensional ◽  
Reliable Model ◽  
Lower Cell Density ◽  
Cell Densities

Three-dimensional in vitro spheroids are a reliable model to study tumor biology and drug toxicity. However, inconsistencies exist in terms of seeding cell density that governs spheroid size and shape, influencing the experimental outcome. We investigated the effect of varying cell densities using glioblastoma cells on tumorsphere formation and their responsiveness to drug treatment. Our results demonstrated that in comparison with spheroids formed with lower cell density, spheroids formed with higher cell density were not only larger in size but also had a larger necrotic core surrounded by a higher number of quiescent cells and were irresponsive to drug treatment. Our study highlights the importance of predetermination of cell density to obtain desired/appropriate spheroid size to produce consistent and reliable data on drug toxicity studies in tumor cells.

Estimating the cell density and invasive radius of three-dimensional glioblastoma tumor spheroids grown in vitro

2007 ◽  
Vol 46 (22) ◽  
pp. 5110 ◽  
Author(s):  
Andrew M. Stein ◽  
Michal O. Nowicki ◽  
Tim Demuth ◽  
Michael E. Berens ◽  
Sean E. Lawler ◽  
...  
Keyword(s):  
Cell Density ◽  
Three Dimensional ◽  
Tumor Spheroids

scholarly journals Promising Applications of Tumor Spheroids and Organoids for Personalized Medicine

Cancers ◽  
2020 ◽  
Vol 12 (10) ◽  
pp. 2727 ◽  
Author(s):  
Zarema Gilazieva ◽  
Aleksei Ponomarev ◽  
Catrin Rutland ◽  
Albert Rizvanov ◽  
Valeriya Solovyeva
Keyword(s):  
Personalized Medicine ◽  
Drug Testing ◽  
Tumor Biology ◽  
Three Dimensional ◽  
3D Models ◽  
Therapeutic Agents ◽  
Therapeutic Strategies ◽  
Tumor Spheroid ◽  
Advantages And Disadvantages

One of the promising directions in personalized medicine is the use of three-dimensional (3D) tumor models such as spheroids and organoids. Spheroids and organoids are three-dimensional cultures of tumor cells that can be obtained from patient tissue and, using high-throughput personalized medicine methods, provide a suitable therapy for that patient. These 3D models can be obtained from most types of tumors, which provides opportunities for the creation of biobanks with appropriate patient materials that can be used to screen drugs and facilitate the development of therapeutic agents. It should be noted that the use of spheroids and organoids would expand the understanding of tumor biology and its microenvironment, help develop new in vitro platforms for drug testing and create new therapeutic strategies. In this review, we discuss 3D tumor spheroid and organoid models, their advantages and disadvantages, and evaluate their promising use in personalized medicine.

scholarly journals 3D bioprinting of high cell-density heterogeneous tissue models through spheroid fusion within self-healing hydrogels

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Andrew C. Daly ◽  
Matthew D. Davidson ◽  
Jason A. Burdick
Keyword(s):  
Cell Density ◽  
Spatial Organization ◽  
High Cell Density ◽  
Self Healing ◽  
High Cell ◽  
Cellular Models ◽  
Functional Features ◽  
Cell Densities ◽  
Tissue Models

AbstractCellular models are needed to study human development and disease in vitro, and to screen drugs for toxicity and efficacy. Current approaches are limited in the engineering of functional tissue models with requisite cell densities and heterogeneity to appropriately model cell and tissue behaviors. Here, we develop a bioprinting approach to transfer spheroids into self-healing support hydrogels at high resolution, which enables their patterning and fusion into high-cell density microtissues of prescribed spatial organization. As an example application, we bioprint induced pluripotent stem cell-derived cardiac microtissue models with spatially controlled cardiomyocyte and fibroblast cell ratios to replicate the structural and functional features of scarred cardiac tissue that arise following myocardial infarction, including reduced contractility and irregular electrical activity. The bioprinted in vitro model is combined with functional readouts to probe how various pro-regenerative microRNA treatment regimes influence tissue regeneration and recovery of function as a result of cardiomyocyte proliferation. This method is useful for a range of biomedical applications, including the development of precision models to mimic diseases and the screening of drugs, particularly where high cell densities and heterogeneity are important.

Self-Assembled β-Sheet Architectures for Bone-Tissue Engineering

1999 ◽  
Vol 599 ◽  
Author(s):  
G. Spreitzer ◽  
J. Doctor ◽  
D. W. Wright
Keyword(s):  
Tissue Engineering ◽  
Bone Tissue Engineering ◽  
Bone Tissue ◽  
Critical Role ◽  
Three Dimensional ◽  
Structural Motif ◽  
Synthetic Approach ◽  
Self Assembled ◽  
Β Sheet

AbstractAdvances in the understanding of biomineralization processes in a variety of organisms have revealed the critical role of three-dimensional scaffolding architectures to create a highly functionalized surface. These complex matrices function on a variety of length scales ranging from the macromolecular (10–100 nm) to the cellular (1–10mm) and larger. One dominant structural motif found in many of these architectures is macromolecules containing antiparallel β-pleated sheets. These “hints” from Nature have lead to the iterative design and development of a novel multipurpose platform technology based on a self-assembled periodic peptide architecture for use in bone-tissue engineering. Combining molecular modeling, structural biochemistry and synthetic techniques, we have produced a β-sheet hollow tube peptide nanoassembly. Such a synthetic approach allows for the template's designed parameters of electrostatic, geometric and stereochemical complimentarily to match those of the desired biomineral. Consequently, these templates readily nucleate calcite. Future studies will investigate the in vitro osteoconductive and osteogenic properties of these templates.

scholarly journals Printing the Pathway Forward in Bone Metastatic Cancer Research: Applications of 3D Engineered Models and Bioprinted Scaffolds to Recapitulate the Bone–Tumor Niche

Cancers ◽  
2021 ◽  
Vol 13 (3) ◽  
pp. 507
Author(s):  
Anne M. Hughes ◽  
Alexus D. Kolb ◽  
Alison B. Shupp ◽  
Kristy M. Shine ◽  
Karen M. Bussard
Keyword(s):  
Cancer Research ◽  
Tumor Cell ◽  
Cancer Cell ◽  
Bone Tumor ◽  
Metastatic Cancer ◽  
Critical Role ◽  
Three Dimensional ◽  
Biocompatible Materials

Breast cancer commonly metastasizes to bone, resulting in osteolytic lesions and poor patient quality of life. The bone extracellular matrix (ECM) plays a critical role in cancer cell metastasis by means of the physical and biochemical cues it provides to support cellular crosstalk. Current two-dimensional in-vitro models lack the spatial and biochemical complexities of the native ECM and do not fully recapitulate crosstalk that occurs between the tumor and endogenous stromal cells. Engineered models such as bone-on-a-chip, extramedullary bone, and bioreactors are presently used to model cellular crosstalk and bone–tumor cell interactions, but fall short of providing a bone-biomimetic microenvironment. Three-dimensional bioprinting allows for the deposition of biocompatible materials and living cells in complex architectures, as well as provides a means to better replicate biological tissue niches in-vitro. In cancer research specifically, 3D constructs have been instrumental in seminal work modeling cancer cell dissemination to bone and bone–tumor cell crosstalk in the skeleton. Furthermore, the use of biocompatible materials, such as hydroxyapatite, allows for printing of bone-like microenvironments with the ability to be implanted and studied in in-vivo animal models. Moreover, the use of bioprinted models could drive the development of novel cancer therapies and drug delivery vehicles.

scholarly journals 3D bioprinting of high cell-density heterogeneous tissue models through spheroid fusion within self-healing hydrogels

2020 ◽  
Author(s):  
Andrew C. Daly ◽  
Matthew D. Davidson ◽  
Jason A. Burdick
Keyword(s):  
Cell Density ◽  
Spatial Organization ◽  
High Cell Density ◽  
Self Healing ◽  
High Cell ◽  
Cellular Models ◽  
Functional Features ◽  
Cell Densities ◽  
Tissue Models

AbstractCellular models are needed to study human development and disease in vitro, including the screening of drugs for toxicity and efficacy. However, current approaches are limited in the engineering of functional tissue models with requisite cell densities and heterogeneity to appropriately model cell and tissue behaviors. Here, we develop a new bioprinting approach to transfer spheroids into self-healing support hydrogels at high resolution, which enables their patterning and fusion into high-cell density microtissues of prescribed spatial organization. As an example application, we bioprint induced pluripotent stem cell-derived cardiac microtissue models with spatially controlled cardiomyocyte and fibroblast cell ratios to replicate the structural and functional features of scarred cardiac tissue that arise following myocardial infarction, including reduced contractility and irregular electrical activity. The bioprinted in vitro model is combined with functional readouts to probe how various pro-regenerative microRNA treatment regimes influence tissue regeneration and recovery of function as a result of cardiomyocyte proliferation. This method is useful for a range of biomedical applications, including the development of precision models to mimic diseases and for the screening of drugs, particularly where high cell densities and heterogeneity are important.

scholarly journals Bi-functional oxidized dextran–based hydrogel inducing microtumors: An in vitro three-dimensional lung tumor model for drug toxicity assays

2017 ◽  
Vol 8 ◽  
pp. 204173141771839 ◽  
Author(s):  
Dhaval Kedaria ◽  
Rajesh Vasita
Keyword(s):  
Drug Screening ◽  
Drug Toxicity ◽  
Disease Modeling ◽  
Lung Tumor ◽  
Three Dimensional ◽  
Two Dimensional ◽  
Chitosan Hydrogel ◽  
Oxidized Dextran

Cancer is a serious death causing disease having 8.2 million deaths in 2012. In the last decade, only about 10% of chemotherapeutic compounds showed productivity in drug screening. Two-dimensional culture assays are the most common in vitro drug screening models, which do not precisely model the in vivo condition for reliable preclinical drug screening. Three-dimensional scaffold–based cell cultures perhaps mimic tumor microenvironment and recapitulate physiologically more relevant tumor. This study was carried out to develop bi-functional oxidized dextran–based cell instructive hydrogel that provides three-dimensional environment to cancer cells for inducing microtumor. Oxidized dextran was blended with thiolated chitosan to fabricate an in situ self-gelable hydrogel (modified dextran–chitosan) in a one-step process. The hydrogels characterization revealed cross-linked network structure with highly porous structure and water absorption. The modified dextran–chitosan hydrogel showed reduced hydrophobicity and has reduced protein absorption, which resulted in changing the A549 cell adhesiveness, and encouraged them to form microtumor. The cells were proliferated in clusters having spherical morphology with randomly oriented stress fiber and large nucleus. Further microtumors were studied for hypoxia where reactive oxygen species generation demonstrated 15-fold increase as compared to monolayer culture. Drug-sensitivity results showed that microtumors generated on modified dextran–chitosan hydrogel showed resistance to doxorubicin with having 33%–58% increased growth than two-dimensional monolayer model at concentrations of 25–100 µM. In summary, the modified dextran–chitosan scaffold can provide surface chemistry that induces three-dimensional microtumors with physiologically relevant properties to in vivo tumor including growth, morphology, extracellular matrix production, hypoxic phenotype, and drug response. This model can be potentially utilized for drug toxicity studies and cancer disease modeling to understand tumor phenotype and progression.

scholarly journals Redirecting differentiation of mammary progenitor cells by 3D bioprinted sweat gland microenvironment

2019 ◽  
Vol 7 ◽  
Author(s):  
Rui Wang ◽  
Yihui Wang ◽  
Bin Yao ◽  
Tian Hu ◽  
Zhao Li ◽  
...  
Keyword(s):  
Progenitor Cells ◽  
Cell Fate ◽  
Sweat Gland ◽  
Critical Role ◽  
Three Dimensional ◽  
Control Group ◽  
Specific Marker ◽  
Shh Pathway

Abstract Background Mammary progenitor cells (MPCs) maintain their reproductive potency through life, and their specific microenvironments exert a deterministic control over these cells. MPCs provides one kind of ideal tools for studying engineered microenvironmental influence because of its accessibility and continually undergoes postnatal developmental changes. The aim of our study is to explore the critical role of the engineered sweat gland (SG) microenvironment in reprogramming MPCs into functional SG cells. Methods We have utilized a three-dimensional (3D) SG microenvironment composed of gelatin-alginate hydrogels and components from mouse SG extracellular matrix (SG-ECM) proteins to reroute the differentiation of MPCs to study the functions of this microenvironment. MPCs were encapsulated into the artificial SG microenvironment and were printed into a 3D cell-laden construct. The expression of specific markers at the protein and gene levels was detected after cultured 14 days. Results Compared with the control group, immunofluorescence and gene expression assay demonstrated that MPCs encapsulated in the bioprinted 3D-SG microenvironment could significantly express the functional marker of mouse SG, sodium/potassium channel protein ATP1a1, and tend to express the specific marker of luminal epithelial cells, keratin-8. When the Shh pathway is inhibited, the expression of SG-associated proteins in MPCs under the same induction environment is significantly reduced. Conclusions Our evidence proved the ability of differentiated mouse MPCs to regenerate SG cells by engineered SG microenvironment in vitro and Shh pathway was found to be correlated with the changes in the differentiation. These results provide insights into regeneration of damaged SG by MPCs and the role of the engineered microenvironment in reprogramming cell fate.

scholarly journals Semiautomatic Growth Analysis of Multicellular Tumor Spheroids

2011 ◽  
Vol 16 (9) ◽  
pp. 1119-1124 ◽  
Author(s):  
Bjoern Rodday ◽  
Franziska Hirschhaeuser ◽  
Stefan Walenta ◽  
Wolfgang Mueller-Klieser
Keyword(s):  
Tumor Biology ◽  
Three Dimensional ◽  
In Vitro Models ◽  
Multicellular Tumor Spheroids ◽  
Tumor Spheroids ◽  
Microtiter Plates ◽  
Accuracy And Precision ◽  
Ocular Micrometer ◽  
Growing Conditions

Multicellular tumor spheroids (MCTS) are routinely employed as three-dimensional in vitro models to study tumor biology. Cultivation of MCTS in spinner flasks provides better growing conditions, especially with regard to the availability of nutrients and oxygen, when compared with microtiter plates. The main endpoint of drug response experiments is spheroid size. It is common practice to analyze spheroid size manually with a microscope and an ocular micrometer. This requires removal of some spheroids from the flask, which entails major limitations such as loss of MCTS and the risk of contamination. With this new approach, the authors present an efficient and highly reproducible method to analyze the size of complete MCTS populations in culture containers with transparent, flat bottoms. MCTS sediments are digitally scanned and spheroid volumes are calculated by computerized image analysis. The equipment includes regular office hardware (personal computer, flatbed scanner) and software (Adobe Photoshop, Microsoft Excel, ImageJ). The accuracy and precision of the method were tested using industrial precision steel beads with known diameter. In summary, in comparison with other methods, this approach provides benefits in terms of semiautomation, noninvasiveness, and low costs.

scholarly journals Cell Stratification, Spheroid Formation and Bioscaffolds Used to Grow Cells in Three Dimensional Cultures

2015 ◽  
Vol 58 (3) ◽  
pp. 79-85 ◽  
Author(s):  
Hana Hrebíková ◽  
Dana Čížková ◽  
Jana Chvátalová ◽  
Rishikaysh Pisal ◽  
Richard Adamčik ◽  
...  
Keyword(s):  
Neural Progenitor Cells ◽  
Three Dimensional ◽  
Semipermeable Membranes ◽  
Novel Drugs ◽  
Invaluable Tool ◽  
In Vitro Systems ◽  
Cell Densities ◽  
And Migration ◽  
Living Tissues

The cell culture became an invaluable tool for studying cell behaviour, development, function, gene expression, toxicity of compounds and efficacy of novel drugs. Although most results were obtained from cell cultivation in two-dimensional (2D) systems, in which cells are grown in a monolayer, three-dimensional (3D) cultures are more promising as they correspond closely to the native arrangement of cells in living tissues. In our study, we focused on three types of 3D in vitro systems used for cultivation of one cell type. Cell morphology, their spatial distribution inside of resulting multicellular structures and changes in time were analysed with histological examination of samples harvested at different time periods. In multilayered cultures of WRL 68 hepatocytes grown on semipermeable membranes and non-passaged neurospheres generated by proliferation of neural progenitor cells, the cells were tightly apposed, showed features of cell differentiation but also cell death that was observable in short-term cultures. Biogenic scaffolds composed of extracellular matrix of the murine tibial anterior muscle were colonized with C2C12 myoblasts in vitro. The recellularized scaffolds did not reach high cell densities comparable with the former systems but supported well cell anchorage and migration without any signs of cell regression.

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