scholarly journals Evaluation of Three-Dimensional Porous Iron-Cross-Linked Alginate as a Scaffold for Cell Culture

2014 ◽  
Vol 2014 ◽  
pp. 1-6 ◽  
Author(s):  
Ikuko Machida-Sano ◽  
Sakito Ogawa ◽  
Makoto Hirakawa ◽  
Hideo Namiki
Keyword(s):  
Cultured Cells ◽  
Porous Scaffold ◽  
Three Dimensional ◽  
Porous Iron ◽  
Multicellular Spheroids ◽  
Cell Scaffolds ◽  
Alginate Gels ◽  
Alginate Scaffold ◽  
Cell Adhesion And Proliferation ◽  
Ca Alginate

We investigated the efficacy of three-dimensional porous ferric-ion-cross-linked alginate (Fe-alginate) gels as cell scaffolds, in comparison with calcium-ion-cross-linked alginate (Ca-alginate) gels. In a previous study, we had demonstrated that two-dimensional Fe-alginate film was an efficient material for use as a scaffold, allowing good cell adhesion and proliferation, unlike Ca-alginate film. In the present study, we fabricated three-dimensional porous Fe- and Ca-alginate gels by freeze-drying and evaluated their effects on cultured cells. The Fe-alginate gels showed higher protein adsorption ability than Ca-alginate gels. Cells formed multicellular spheroids in both types of alginate scaffold, but the number of cultured cells increased with culture time on Fe-alginate porous gels, whereas those on Ca-alginate gels did not. Moreover, it was revealed that the cells on Fe-alginate scaffolds were still viable inside the multicellular spheroids even after cultivation for 14 days. These results suggest that Fe-alginate provides a superior porous scaffold suitable for three-dimensional culture of cells. Our findings may be useful for extending the application of Fe-alginate to diverse biomedical fields.

scholarly journals Copper-64 Chloride Exhibits Therapeutic Potential in Three-Dimensional Cellular Models of Prostate Cancer

2020 ◽  
Vol 7 ◽  
Author(s):  
Catarina I. G. Pinto ◽  
Sara Bucar ◽  
Vítor Alves ◽  
Alexandra Fonseca ◽  
Antero J. Abrunhosa ◽  
...  
Keyword(s):  
Prostate Cancer ◽  
Cultured Cells ◽  
Therapeutic Potential ◽  
Three Dimensional ◽  
Diagnostic Methods ◽  
Cancer Type ◽  
Cancer Resistance ◽  
Multicellular Spheroids ◽  
Cellular Models ◽  
Theranostic Agent

Prostate cancer (PCa) is the second most common cancer type in men, and in advanced metastatic stages is considerable incurable. This justifies the need for efficient early diagnostic methods and novel therapies, particularly radiopharmaceuticals with the potential for simultaneous diagnosis and therapy (theranostics). We have previously demonstrated, using monolayer-cultured cells, that copper-64 chloride, a promising theranostic agent for PCa, has the potential to induce significant damage in cancer cells while having minimal side effects in healthy tissues. Here, we further explored this compound for its theranostic applications using more advanced PCa cellular models, specifically multicellular spheroids. Namely, we evaluated the cellular uptake of 64CuCl2 in three human PCa spheroids (derived from 22RV1, DU145, and LNCaP cells), and characterized the growth profile and viability of those spheroids as well as the clonogenic capacity of spheroid-derived cells after exposure to 64CuCl2. Furthermore, the populations of cancer stem cells (CSCs), known to be important for cancer resistance and recurrence, present in the spheroid models were also evaluated using two different markers (CD44 and CD117). 64CuCl2 was found to have significant detrimental effects in spheroids and spheroid-derived cells, being able to reduce their growth and impair the viability and reproductive ability of spheroids from both castration-resistant (22RV1 and DU145) and hormone-naïve PCa (LNCaP). Interestingly, resistance to 64CuCl2 treatment seemed to be related with the presence of a CSC population, since the most resistant spheroids, derived from the DU145 cell line, had the highest initial percentage of CSCs among the three cell lines under study. Altogether, these results clearly highlight the theranostic potential of 64CuCl2.

Three-dimensional Growth of Renal Epithelial Cells in Vitro: A Tool in Toxicity Testing

1993 ◽  
Vol 21 (2) ◽  
pp. 191-195 ◽  
Author(s):  
Knut-Jan Andersen ◽  
Erik Ilsø Christensen ◽  
Hogne Vik
Keyword(s):  
Epithelial Cells ◽  
Three Dimensional ◽  
Toxicity Testing ◽  
Renal Tissue ◽  
Epithelial Cell Line ◽  
Multicellular Spheroids ◽  
Renal Epithelial Cell ◽  
Renal Epithelial Cells

The tissue culture of multicellular spheroids from the renal epithelial cell line LLC-PK1 (proximal tubule) is described. This represents a biological system of intermediate complexity between renal tissue in vivo and simple monolayer cultures. The multicellular structures, which show many similarities to kidney tubules in vivo, including a vectorial water transport, should prove useful for studying the potential nephrotoxicity of drugs and chemicals in vitro. In addition, the propagation of renal epithelial cells as multicellular spheroids in serum-free culture may provide information on the release of specific biological parameters, which may be suppressed or masked in serum-supplemented media.

scholarly journals Reversal of senescence-associated beta-galactosidase expression during in vitro three-dimensional tissue-engineering of human chondrocytes in a polymer scaffold

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Shojiro Katoh ◽  
Atsuki Fujimaru ◽  
Masaru Iwasaki ◽  
Hiroshi Yoshioka ◽  
Rajappa Senthilkumar ◽  
...  
Keyword(s):  
Regenerative Medicine ◽  
Replicative Senescence ◽  
Cultured Cells ◽  
Three Dimensional ◽  
Human Chondrocytes ◽  
Cartilage Tissue ◽  
Optimal Method ◽  
Beta Galactosidase

AbstractRegenerative medicine applications require cells that are not inflicted with senescence after in vitro culture for an optimal in vivo outcome. Methods to overcome replicative senescence include genomic modifications which have their own disadvantages. We have evaluated a three-dimensional (3D) thermo-reversible gelation polymer (TGP) matrix environment for its capabilities to reverse cellular senescence. The expression of senescence-associated beta-galactosidase (SA-βgal) by human chondrocytes from osteoarthritis-affected cartilage tissue, grown in a conventional two-dimensional (2D) monolayer culture versus in 3D-TGP were compared. In 2D, the cells de-differentiated into fibroblasts, expressed higher SA-βgal and started degenerating at 25 days. SA-βgal levels decreased when the chondrocytes were transferred from the 2D to the 3D-TGP culture, with cells exhibiting a tissue-like growth until 42–45 days. Other senescence associated markers such as p16INK4a and p21 were also expressed only in 2D cultured cells but not in 3D-TGP tissue engineered cartilage. This is a first-of-its-kind report of a chemically synthesized and reproducible in vitro environment yielding an advantageous reversal of aging of human chondrocytes without any genomic modifications. The method is worth consideration as an optimal method for growing cells for regenerative medicine applications.

scholarly journals Porous Geometry Guided Micro-mechanical Environment Within Scaffolds for Cell Mechanobiology Study in Bone Tissue Engineering

2021 ◽  
Vol 9 ◽  
Author(s):  
Feihu Zhao ◽  
Yi Xiong ◽  
Keita Ito ◽  
Bert van Rietbergen ◽  
Sandra Hofmann
Keyword(s):  
Porous Scaffold ◽  
Mechanical Strain ◽  
Three Dimensional ◽  
Pore Geometry ◽  
Cell Physiology ◽  
Porous Scaffolds ◽  
Mechanical Environment ◽  
Functional Features ◽  
Future Work

Mechanobiology research is for understanding the role of mechanics in cell physiology and pathology. It will have implications for studying bone physiology and pathology and to guide the strategy for regenerating both the structural and functional features of bone. Mechanobiological studies in vitro apply a dynamic micro-mechanical environment to cells via bioreactors. Porous scaffolds are commonly used for housing the cells in a three-dimensional (3D) culturing environment. Such scaffolds usually have different pore geometries (e.g. with different pore shapes, pore dimensions and porosities). These pore geometries can affect the internal micro-mechanical environment that the cells experience when loaded in the bioreactor. Therefore, to adjust the applied micro-mechanical environment on cells, researchers can tune either the applied load and/or the design of the scaffold pore geometries. This review will provide information on how the micro-mechanical environment (e.g. fluid-induced wall shear stress and mechanical strain) is affected by various scaffold pore geometries within different bioreactors. It shall allow researchers to estimate/quantify the micro-mechanical environment according to the already known pore geometry information, or to find a suitable pore geometry according to the desirable micro-mechanical environment to be applied. Finally, as future work, artificial intelligent – assisted techniques, which can achieve an automatic design of solid porous scaffold geometry for tuning/optimising the micro-mechanical environment are suggested.

scholarly journals A Matrigel-based 3D construct of SH-SY5Y cells models the α-synuclein pathologies of Parkinson's disease

2022 ◽  
Author(s):  
Zhao-Feng Li ◽  
Lei Cui ◽  
Mi-Mi Jin ◽  
Dong-Yan Hu ◽  
Xiao-Gang Hou ◽  
...  
Keyword(s):  
Parkinson’S Disease ◽  
Parkinson's Disease ◽  
Lewy Body ◽  
Cultured Cells ◽  
Three Dimensional ◽  
Proteinase K ◽  
All Trans Retinoic Acid ◽  
Lewy Body Pathology ◽  
Rotenone Treatment ◽  
2D And 3D

Parkinson's disease (PD) is featured with α-synuclein-based Lewy body pathology, which however was difficult to observe in conventional two-dimensional (2D) cell culture and even in animal models. We herein aimed to develop a three-dimensional (3D) cellular model of PD to recapitulate the α-synuclein pathologies. All-trans-retinoic acid-differentiated human SH-SY5Y cells and Matrigel were optimized for 3D construction. The 3D cultured cells displayed higher tyrosine hydroxylase expression and improved dopaminergic-like phenotypes than 2D cells as suggested by RNA-sequencing analyses. Multiple forms of α-synuclein, including monomer, low and high molecular weight oligomers, were differentially present in the 2D and 3D cells, but mostly remained unchanged upon the MPP+ or rotenone treatment. Phosphorylated α-synuclein was accumulated and detergent-insoluble α-synuclein fraction was observed in the neurotoxin-treated 3D cells. Importantly, Lewy body-like inclusions were captured in the 3D system, including proteinase K-resistant α-synuclein aggregates, ubiquitin aggregation, β-amyloid and β-sheet protein deposition. The study provides a unique and convenient 3D model of PD which recapitulates critical α-synuclein pathologies and should be useful in multiple PD-associated applications.

scholarly journals Using Spheroids as Building Blocks Towards 3D Bioprinting of Tumor Microenvironment

2021 ◽  
Vol 7 (4) ◽  
pp. 444
Author(s):  
Pei Zhuang ◽  
Yi-Hua Chiang ◽  
Maria Serafim Fernanda ◽  
Mei He
Keyword(s):  
Tumor Microenvironment ◽  
Prediction Models ◽  
Three Dimensional ◽  
3D Culture ◽  
Building Blocks ◽  
3D Bioprinting ◽  
Multicellular Spheroids ◽  
Tumor Models ◽  
3D Printed

Cancer still ranks as a leading cause of mortality worldwide. Although considerable efforts have been dedicated to anticancer therapeutics, progress is still slow, partially due to the absence of robust prediction models. Multicellular tumor spheroids, as a major three-dimensional (3D) culture model exhibiting features of avascular tumors, gained great popularity in pathophysiological studies and high throughput drug screening. However, limited control over cellular and structural organization is still the key challenge in achieving in vivo like tissue microenvironment. 3D bioprinting has made great strides toward tissue/organ mimicry, due to its outstanding spatial control through combining both cells and materials, scalability, and reproducibility. Prospectively, harnessing the power from both 3D bioprinting and multicellular spheroids would likely generate more faithful tumor models and advance our understanding on the mechanism of tumor progression. In this review, the emerging concept on using spheroids as a building block in 3D bioprinting for tumor modeling is illustrated. We begin by describing the context of the tumor microenvironment, followed by an introduction of various methodologies for tumor spheroid formation, with their specific merits and drawbacks. Thereafter, we present an overview of existing 3D printed tumor models using spheroids as a focus. We provide a compilation of the contemporary literature sources and summarize the overall advancements in technology and possibilities of using spheroids as building blocks in 3D printed tissue modeling, with a particular emphasis on tumor models. Future outlooks about the wonderous advancements of integrated 3D spheroidal printing conclude this review.

scholarly journals An efficient polymer cocktail-based transportation method for cartilage tissue, yielding chondrocytes with enhanced hyaline cartilage expression during in vitro culturing

2021 ◽  
Author(s):  
Shojiro Katoh ◽  
Hiroshi Yoshioka ◽  
Shoji Suzuki ◽  
Hiroyuki Nakajima ◽  
Masaru Iwasaki ◽  
...  
Keyword(s):  
Cultured Cells ◽  
Hyaline Cartilage ◽  
Three Dimensional ◽  
Cartilage Tissue ◽  
Human Cartilage ◽  
Tissue Samples ◽  
Chondrocyte Implantation ◽  
Initial Processing ◽  
Regenerative Therapies

Chondrocytes are used in cell-based therapies such as autologous chondrocyte implantation (ACI) and matrix-associated cartilage implantation (MACI). To transport the cartilage tissue to the laboratory for in vitro culturing, phosphate-buffered saline (PBS), Euro-Collins solution (ECS) and Dulbecco Modified Eagle Medium (DMEM) are commonly employed at 4-8 deg C. In this study, eight samples of human cartilage biopsy tissues from elderly patients with severe osteoarthritis undergoing arthroscopy, which would otherwise have been discarded, were used. The cartilage tissue samples were compared to assess the cell yield between two transportation groups: i) a thermo-reversible gelation polymer (TGP) based method without cool preservation (~25 deg C) and ii) ECS transport at 4 deg C. These samples were subjected to in vitro culture in a two-dimensional (2D) monolayer for two weeks and subsequently in a three-dimensional (3D) TGP scaffold for six weeks. The cell count obtained from the tissues transported in TGP was higher (0.2 million cells) than those transported in ECS (0.08 million cells) both after initial processing and after in vitro culturing for 2 weeks in 2D (18 million cells compared with 10 million cells). In addition, mRNA quantification demonstrated significantly higher expression of Col2a1 and SOX-9 in 3D-TGP cultured cells and lower expression of COL1a1 in RT-PCR, characteristic of the hyaline cartilage phenotype, than in 2D culture. This study confirms that the TGP cocktail is suitable for both the transport of human cartilage tissue and for in vitro culturing to yield better-quality cells for use in regenerative therapies.

Development of Scaffold Building Units and Assembly for Tissue Engineering Using Fused Deposition Modelling

2009 ◽  
Vol 83-86 ◽  
pp. 269-274 ◽  
Author(s):  
Syed H. Masood ◽  
Kadhim Alamara
Keyword(s):  
Tissue Engineering ◽  
Pore Size ◽  
Fused Deposition Modeling ◽  
Porous Scaffold ◽  
Cell Structure ◽  
Three Dimensional ◽  
Three Dimensions ◽  
Fused Deposition Modelling ◽  
Fused Deposition ◽  
Unit Cells

In tissue engineering (TE), a porous scaffold structure of biodegradable material is required as a template to guide the proliferation, growth and development of cells appropriately in three dimensions. The scaffold must meet design requirements of appropriate porosity, pore size and interconnected structure to allow cell proliferation and adhesion. This paper presents a methodology for design and manufacture of TE scaffolds with varying porosity by employing open structure building units and Fused Deposition Modeling (FDM) rapid prototyping technique. A computer modeling approach for constructing and assembly of three-dimensional unit cell structure is presented to provide a solution of scaffolds design that can potentially meet the diverse requirements of TE applications. A parametric set of open polyhedral unit cells is used to assist the user in designing the required micro-architecture of the scaffold with required porosity and pore size and then the Boolean operation is used to create the scaffold of a given CAD model from the designed microstructure. The procedure is verified by fabrication of physical scaffolds using the commercial FDM system.

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