scholarly journals Hybrid multi-layered scaffolds produced via grain extrusion and electrospinning for 3D cell culture tests

2019 ◽  
Vol 26 (3) ◽  
pp. 593-602
Author(s):  
Paola Ginestra ◽  
Stefano Pandini ◽  
Elisabetta Ceretti
Keyword(s):  
Cell Attachment ◽  
Three Dimensional ◽  
3D Culture ◽  
3D Cell Culture ◽  
Optical Microscope ◽  
Biological Interactions ◽  
Compression Tests ◽  
Content Type ◽  
Extrusion Head ◽  
Mechanical Performances

Purpose The purpose of this paper is to focus on the production of scaffolds with specific morphology and mechanical behavior to satisfy specific requirements regarding their stiffness, biological interactions and surface structure that can promote cell-cell and cell-matrix interactions though proper porosity, pore size and interconnectivity. Design/methodology/approach This case study was focused on the production of multi-layered hybrid scaffolds made of polycaprolactone and consisting in supporting grids obtained by Material Extrusion (ME) alternated with electrospun layers. An open source 3D printer was utilized, with a grain extrusion head that allows the production and distribution of strands on the plate according to the designed geometry. Square grid samples were observed under optical microscope showing a good interconnectivity and spatial distribution of the pores, while scanning electron microscope analysis was used to study the electrospun mats morphology. Findings A good adhesion between the ME and electrospinning layers was achieved by compression under specific thermomechanical conditions obtaining a hybrid three-dimensional scaffold. The mechanical performances of the scaffolds have been analyzed by compression tests, and the biological characterization was carried out by seeding two different cells phenotypes on each side of the substrates. Originality/value The structure of the multi-layered scaffolds demonstrated to play an important role in promoting cell attachment and proliferation in a 3D culture formation. It is expected that this design will improve the performances of osteochondral scaffolds with a strong influence on the required formation of an interface tissue and structure that need to be rebuilt.

scholarly journals Applications of Biomaterials in 3D Cell Culture and Contributions of 3D Cell Culture to Drug Development and Basic Biomedical Research

2021 ◽  
Vol 22 (5) ◽  
pp. 2491
Author(s):  
Yujin Park ◽  
Kang Moo Huh ◽  
Sun-Woong Kang
Keyword(s):  
Cell Culture ◽  
Three Dimensional ◽  
3D Culture ◽  
3D Cell Culture ◽  
Accurate Method ◽  
New Drugs ◽  
Toxicity Screening ◽  
Cellular Functions ◽  
Toxicity Of Drugs ◽  
External Stimuli

The process of evaluating the efficacy and toxicity of drugs is important in the production of new drugs to treat diseases. Testing in humans is the most accurate method, but there are technical and ethical limitations. To overcome these limitations, various models have been developed in which responses to various external stimuli can be observed to help guide future trials. In particular, three-dimensional (3D) cell culture has a great advantage in simulating the physical and biological functions of tissues in the human body. This article reviews the biomaterials currently used to improve cellular functions in 3D culture and the contributions of 3D culture to cancer research, stem cell culture and drug and toxicity screening.

A rhabdomyosarcoma hydrogel model to unveil cell-extracellular matrix interactions

2021 ◽  
Author(s):  
Mattia Saggioro ◽  
Stefania D'Agostino ◽  
Anna Gallo ◽  
Sara Crotti ◽  
Sara D'Aronco ◽  
...  
Keyword(s):  
Cell Culture ◽  
Extracellular Matrix ◽  
Three Dimensional ◽  
3D Culture ◽  
3D Cell Culture ◽  
Culture Systems ◽  
Matrix Interactions ◽  
In Vitro Systems ◽  
Extracellular Matrix Interactions

Three-dimensional (3D) culture systems are progressively getting attention given their potential in overcoming limitations of the classical 2D in vitro systems. Among different supports for 3D cell culture, hydrogels (HGs)...

scholarly journals 3D cell culture technology – a new insight into in vitro research – a review

2021 ◽  
Vol 0 (0) ◽  
Author(s):  
Justyna Sośniak ◽  
Jolanta Opiela
Keyword(s):  
Three Dimensional ◽  
3D Culture ◽  
3D Cell Culture ◽  
2D Systems ◽  
Multicellular Spheroids ◽  
Advantages And Disadvantages ◽  
3D Cell Cultures ◽  
Culture Models ◽  
Culture Technology

Abstract Most in vitro cell-based research is based on two-dimensional (2D) systems where growth and development take place on a flat surface, which does not reflect the natural environment of the cells. The imperfection and limitations of culture in 2D systems eventually led to the creation of three-dimensional (3D) culture models that more closely reproduce the actual conditions of physiological cell growth. Since the inception of 3D culture technology, many culture models have been developed, such as technologies of multicellular spheroids, organoids, and organs on chips in the technology of scaffolding, hydrogels, bio-printing and liquid media. In this review we will focus on the advantages and disadvantages of the 2D vs. 3D cell cultures technologies. We will also try to sum up available 3D cultures systems and materials for building 3D scaffolds.

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.

A three-dimensional collagen matrix as a suitable culture system for the comparison of cyclic strain and hydrostatic pressure effects on intervertebral disc cells

2005 ◽  
Vol 2 (4) ◽  
pp. 457-465 ◽  
Author(s):  
Cornelia Neidlinger-Wilke ◽  
Karin Würtz ◽  
Astrid Liedert ◽  
Carla Schmidt ◽  
Wolfgang Börm ◽  
...  
Keyword(s):  
Hydrostatic Pressure ◽  
Intervertebral Disc ◽  
Three Dimensional ◽  
3D Culture ◽  
Cyclic Strain ◽  
Collagen Matrix ◽  
Disc Cell ◽  
Content Type ◽  
Disc Cells ◽  
Fine Print

Object. To study intervertebral disc cell mechanobiology, the authors developed experimental systems that allow the application of cyclic strain and intermittent hydrostatic pressure (IHP) on isolated disc cells under equal three-dimensional (3D) culture conditions. The purpose of the study was to characterize disc cell proliferation, viability, morphology, and gene expression in 3D collagen matrices. Methods. The effects of cyclic strain (1, 2, 4, and 8% strain; 1 Hz) and IHP (0.25 MPa, 0.1 Hz) on gene expression (real-time polymerase chain reaction) of anabolic and catabolic matrix proteins were investigated and compared with those derived from mechanically unstimulated controls. Intervertebral disc cells proliferated in the collagen gels (mean viability 91.6%) and expressed messenger RNA for collagen I, collagen II, aggrecan, matrix metalloproteinase (MMP)—2, and MMP-3. Morphologically, both spindle-shaped cells with longer processes and rounded cells were detected in the collagen scaffolds. Cyclic strain increased collagen II and aggrecan expression and decreased MMP-3 expression of anulus fibrosus cells. No significant difference between the four strain magnitudes was found. Intermittent hydrostatic pressure tended to increase collagen I and aggrecan expression of nucleus cells and significantly decreased MMP-2 and -3 expression of nucleus cells, whereas aggrecan expression of anulus cells tended to decrease. Conclusions. Based on these results, the collagen matrix appeared to be a suitable substrate to apply both cyclic strain and IHP to intervertebral disc cells under 3D culture conditions. Individual variations may be influenced by the extent of degeneration of the disc specimens from which the cells were isolated. This experimental setup may be suitable for studying the influence of degeneration on the disc cell response to mechanical stimuli.

Fabrication and In-Vitro Characterization of Three-Dimensional Composite Scaffolds by Robocasting for Biomedical Applications

2006 ◽  
Vol 49 ◽  
pp. 153-158 ◽  
Author(s):  
J. Russias ◽  
Eduardo Saiz ◽  
Sylvain Deville ◽  
Antoni P. Tomsia
Keyword(s):  
Mechanical Response ◽  
Methylene Chloride ◽  
Cell Attachment ◽  
Three Dimensional ◽  
Inorganic Materials ◽  
Cartilage Defects ◽  
Plastic Behavior ◽  
Compression Tests ◽  
Freeform Fabrication

The development of novel biodegradable scaffolds for the treatment of bone and cartilage defects is the subject of intense research. A successful scaffold will guide cell-attachment, proliferation and tissue regeneration. The objective of this study is to use freeform fabrication (robocasting) for the preparation of porous hybrid organic/inorganic materials with a well controlled architecture and porosity. Polymer/hydroxyapatite (HA) pastes with ceramic contents ranging between 0 to 70 wt. % are prepared by mixing ceramic powders with a solution of the polymer in methylene chloride. Two different polymers are studied: polylactide (PLA) and polycaprolactone (PCL). During the compression tests, the scaffolds show an elasto-plastic behavior with large plastic yielding and do not fail in a brittle manner. The mechanical response is anisotropic and depends significantly on the ceramic content and the type of polymer.

scholarly journals Influence of electrohydrodynamic jetting parameters on the morphology of PCL scaffolds

2017 ◽  
Vol 3 (1) ◽  
Author(s):  
Hang Liu ◽  
Sanjairaj Vijayavenkataraman ◽  
Dandan Wang ◽  
Linzhi Jing ◽  
Jie Sun ◽  
...  
Keyword(s):  
Cell Culture ◽  
Cell Attachment ◽  
3D Cell Culture ◽  
Solution Concentration ◽  
Processing Parameters ◽  
Optical Microscope ◽  
3D Scaffold ◽  
Sem Images ◽  
Scaffold Fabrication ◽  
Fibre Characteristics

 One of the important constituents in tissue engineering is scaffold, which provides structural support and suitable microenvironment for the cell attachment, growth and proliferation. To fabricate micro/nano structures for soft tissue repair and three-dimensional (3D) cell culture, the key is to improve fibre-based scaffold fabrication. Electrohydrodynamic (EHD) jetting is capable of producing and orientating submicron fibres for 3D scaffold fabrication. In this work, an EHD-jetting system was developed to explore the relationship between vital processing parameters and fibre characteristics. In this study, polycaprolactone (PCL) solution prepared by dissolving PCL pellets in acetic acid was used to fabricate the scaffolds. The influence of voltage, motorized stage speed, solution feed rate, and solution concentration on fibre characteristics and scaffold pattern were studied. Morphology of the EHD-jetted PCL fibres and scaffolds were analysed using optical microscope images and scanning electron microscope (SEM) images. Multi-layer scaffolds with the varied coiled pattern were fabricated and analysed. Cell attachment and proliferation have to be investigated in the future by further cell culture studies on these multi-layer coiled scaffolds.

scholarly journals Up-regulated fibronectin in 3D culture facilitates spreading of triple negative breast cancer cells on 2D through integrin β-5 and Src

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Hwa -Jeong Park ◽  
David M. Helfman
Keyword(s):  
Breast Cancer ◽  
Cancer Cells ◽  
Triple Negative Breast Cancer ◽  
Triple Negative ◽  
Cell Attachment ◽  
Three Dimensional ◽  
3D Culture ◽  
Cell Spreading ◽  
Mitogen Activated Protein Kinase ◽  
Rgd Peptides

AbstractUsing MDA-MB-231 cells as a model of triple negative breast cancer (TNBC) and its metastatic sub-cell lines that preferentially metastasize to lung, bone or brain, we found that the mRNA and protein levels of fibronectin (FN) are increased in MDA-MB-231 cells and its lung metastatic derivative, when cultivated in three-dimensional (3D) suspension cultures. The increase of FN expression in 3D was dependent on p38 mitogen-activated protein kinase (MAPK) because it was prevented by treatment of cells with SB203580, an inhibitor of p38MAPK. The up-regulated FN was converted into fibrils, and it enhanced cell spreading when cells cultured in 3D were transferred to two-dimensional (2D) culture. The arginine-glycine-aspartate (RGD) peptides and siRNAs targeting of integrin β-5 inhibited spreading of cells regardless of the presence of FN on 2D culture dishes. In addition, the levels of phosphorylated Src were found to be increased in 3D and the treatment of cells with SU6656, an inhibitor of Src, decreased the rate of cell spreading on FN. Collectively, these studies demonstrate that increased cellular FN in 3D suspension culture facilitates cancer cell attachment and spreading via integrin β-5 and Src, suggesting that the increased FN promotes initial attachment of cancer cells to secondary organs after circulation during metastasis.

scholarly journals A Microfluidic Chip Embracing a Nanofiber Scaffold for 3D Cell Culture and Real-Time Monitoring

Nanomaterials ◽  
2019 ◽  
Vol 9 (4) ◽  
pp. 588 ◽  
Author(s):  
Jeong Hwa Kim ◽  
Ju Young Park ◽  
Songwan Jin ◽  
Sik Yoon ◽  
Jong-Young Kwak ◽  
...  
Keyword(s):  
Cell Culture ◽  
Real Time ◽  
Microfluidic Chip ◽  
Three Dimensional ◽  
3D Culture ◽  
3D Cell Culture ◽  
Culture Conditions ◽  
Real Time Monitoring ◽  
Nanofiber Scaffold

Recently, three-dimensional (3D) cell culture and tissue-on-a-chip application have attracted attention because of increasing demand from the industries and their potential to replace conventional two-dimensional culture and animal tests. As a result, numerous studies on 3D in-vitro cell culture and microfluidic chip have been conducted. In this study, a microfluidic chip embracing a nanofiber scaffold is presented. A electrospun nanofiber scaffold can provide 3D cell culture conditions to a microfluidic chip environment, and its perfusion method in the chip can allow real-time monitoring of cell status based on the conditioned culture medium. To justify the applicability of the developed chip to 3D cell culture and real-time monitoring, HepG2 cells were cultured in the chip for 14 days. Results demonstrated that the cells were successfully cultured with 3D culture-specific-morphology in the chip, and their albumin and alpha-fetoprotein production was monitored in real-time for 14 days.

scholarly journals High-throughput 3D screening for differentiation of hPSC-derived cell therapy candidates

2020 ◽  
Vol 6 (32) ◽  
pp. eaaz1457
Author(s):  
Riya Muckom ◽  
Xiaoping Bao ◽  
Eric Tran ◽  
Evelyn Chen ◽  
Abirami Murugappan ◽  
...  
Keyword(s):  
Cell Therapy ◽  
High Throughput ◽  
System Development ◽  
Three Dimensional ◽  
3D Culture ◽  
3D Cell Culture ◽  
Culture Conditions ◽  
Nervous System Development ◽  
Oligodendrocyte Progenitor Cells ◽  
Cell Therapies

The emergence of several cell therapy candidates in the clinic is an encouraging sign for human diseases/disorders that currently have no effective treatment; however, scalable production of these cell therapies has become a bottleneck. To overcome this barrier, three-dimensional (3D) cell culture strategies have been considered for enhanced cell production. Here, we demonstrate a high-throughput 3D culture platform used to systematically screen 1200 culture conditions with varying doses, durations, dynamics, and combinations of signaling cues to derive oligodendrocyte progenitor cells and midbrain dopaminergic neurons from human pluripotent stem cells (hPSCs). Statistical models of the robust dataset reveal previously unidentified patterns about cell competence to Wnt, retinoic acid, and sonic hedgehog signals, and their interactions, which may offer insights into the combinatorial roles these signals play in human central nervous system development. These insights can be harnessed to optimize production of hPSC-derived cell replacement therapies for a range of neurological indications.

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