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Biomedicines
Special Issues
3D Cell and Tissue Culture
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3D Cell and Tissue Culture

A special issue of Biomedicines (ISSN 2227-9059). This special issue belongs to the section "Molecular and Translational Medicine".

Deadline for manuscript submissions: closed (31 May 2020) | Viewed by 26764

Special Issue Editor

Dr. Matthias Nees

E-Mail Website
Guest Editor
Department of Biochemistry and Molecular Biology, Medical University of Lublin, 20-093 Lublin, Poland
Interests: tumor microenvironment (TME); extracellular matrix (ECM); tissue engineering; organotypic models; microfluidics; organ-on-chip; patient-derived organoids (PDOs); patient-derived explants (PDEs); tumor/stroma interaction; desmoplastic stroma
Special Issues, Collections and Topics in MDPI journals

Special Issue Information

Dear Colleagues,

Three-dimensional cell and tissue cultures have been around for some time, but are still considered as emerging technologies. This is due to the rapidly evolving methods for a growing scope of very diverse applications. In this Special Issue of Biomedicines, it is our goal to present outstanding publications that represent the cutting edge(s) of this development. In principle, all 3D cultures aim for a more faithful and biologically relevant recapitulation of tissue functions. Accordingly, 3D cultures range from relatively simple cell aggregates (spheroids) to increasingly functional organ-like or organotypic microtissues (organoids and tissue-on-a-chip).

Contributions are invited that focus on five core areas of 3D cell and tissue cultures:

  • High-content screening: The implementation of 3D cell and tissue cultures (usually spheroids) for early stage drug and lead discovery, primarily used in pharmaceutical research.
  • Organotypic cultures: Aiming for the formation of complex structures and tissue architectures in vitro. Typically used for the functional validation of biomarkers and (drug) targets or chemosensitivity testing. Many applications include key functional aspects such as extracellular matrix and the microenvironment, or co-cultures of several cell types.
  • Automated imaging and image analysis tools: Aiming for the quantitative assessment of phenotypic features observed in 3D cultures. Complex solutions often require machine vision and artificial intelligence applications.
  • Organoids and tissue cultures: These include slices, explant cultures, etc. for personalized medicine and chemosensitivity testing, aiming at the improvement of personalized medicine and clinical applications.
  • Organ-on-a-chip applications: These cover the most diverse and complex goals—typically related to the recapitulation of biological organ function or failure, tissue formation, and disease mechanisms.

This Special Issue welcomes both original papers and review articles addressing one or several of the above issues, as well as related topics not explicitly mentioned. Overall, we hope that this timely focused Issue summarizing our current knowledge on three-dimensional cell and tissue cultures will be of interest to a wide range of researchers in this field.

Dr. Matthias Nees
Guest Editor

Manuscript Submission Information

Manuscripts should be submitted online at www.mdpi.com by registering and logging in to this website. Once you are registered, click here to go to the submission form. Manuscripts can be submitted until the deadline. All submissions that pass pre-check are peer-reviewed. Accepted papers will be published continuously in the journal (as soon as accepted) and will be listed together on the special issue website. Research articles, review articles as well as short communications are invited. For planned papers, a title and short abstract (about 100 words) can be sent to the Editorial Office for announcement on this website.

Submitted manuscripts should not have been published previously, nor be under consideration for publication elsewhere (except conference proceedings papers). All manuscripts are thoroughly refereed through a single-blind peer-review process. A guide for authors and other relevant information for submission of manuscripts is available on the Instructions for Authors page. Biomedicines is an international peer-reviewed open access monthly journal published by MDPI.

Please visit the Instructions for Authors page before submitting a manuscript. The Article Processing Charge (APC) for publication in this open access journal is 2600 CHF (Swiss Francs). Submitted papers should be well formatted and use good English. Authors may use MDPI's English editing service prior to publication or during author revisions.

Keywords

  • organotypic cultures
  • organoids
  • spheroids
  • machine vision
  • automated image analyses
  • phenotypic screening
  • high-content screening (HCS)
  • organ-on-a-chip
  • tissue culture
  • tissue slices and tissue explants
  • tissue recombination
  • de-cellularized tissues

Published Papers (5 papers)

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Research

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19 pages, 4344 KiB  
Article
Vasculogenesis from Human Dental Pulp Stem Cells Grown in Matrigel with Fully Defined Serum-Free Culture Media
by Jon Luzuriaga, Jon Irurzun, Igor Irastorza, Fernando Unda, Gaskon Ibarretxe and Jose R. Pineda
Biomedicines 2020, 8(11), 483; https://doi.org/10.3390/biomedicines8110483 - 09 Nov 2020
Cited by 27 | Viewed by 4114
Abstract
The generation of vasculature is one of the most important challenges in tissue engineering and regeneration. Human dental pulp stem cells (hDPSCs) are some of the most promising stem cell types to induce vasculogenesis and angiogenesis as they not only secrete vascular endothelial [...] Read more.
The generation of vasculature is one of the most important challenges in tissue engineering and regeneration. Human dental pulp stem cells (hDPSCs) are some of the most promising stem cell types to induce vasculogenesis and angiogenesis as they not only secrete vascular endothelial growth factor (VEGF) but can also differentiate in vitro into both endotheliocytes and pericytes in serum-free culture media. Moreover, hDPSCs can generate complete blood vessels containing both endothelial and mural layers in vivo, upon transplantation into the adult brain. However, many of the serum free media employed for the growth of hDPSCs contain supplements of an undisclosed composition. This generates uncertainty as to which of its precise components are necessary and which are dispensable for the vascular differentiation of hDPSCs, and also hinders the transfer of basic research findings to clinical cell therapy. In this work, we designed and tested new endothelial differentiation media with a fully defined composition using standard basal culture media supplemented with a mixture of B27, heparin and growth factors, including VEGF-A165 at different concentrations. We also optimized an in vitro Matrigel assay to characterize both the ability of hDPSCs to differentiate to vascular cells and their capacity to generate vascular tubules in 3D cultures. The description of a fully defined serum-free culture medium for the induction of vasculogenesis using human adult stem cells highlights its potential as a relevant innovation for tissue engineering applications. In conclusion, we achieved efficient vasculogenesis starting from hDPSCs using serum-free culture media with a fully defined composition, which is applicable for human cell therapy purposes. Full article
(This article belongs to the Special Issue 3D Cell and Tissue Culture)
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13 pages, 3259 KiB  
Article
Image Analysis of 3D Conjunctival Melanoma Cell Cultures Following Electrochemotherapy
by Miltiadis Fiorentzis, Periklis Katopodis, Helen Kalirai, Berthold Seitz, Arne Viestenz and Sarah E. Coupland
Biomedicines 2020, 8(6), 158; https://doi.org/10.3390/biomedicines8060158 - 13 Jun 2020
Cited by 5 | Viewed by 3289
Abstract
Three-dimensional (3D) cell cultures represent small avascular tumors in vitro and simulate some of the biological characteristics of solid tumors, enhancing the evaluation of anticancer drug efficacy. Automated image analysis can be used for the assessment of tumor growth and documentation of changes [...] Read more.
Three-dimensional (3D) cell cultures represent small avascular tumors in vitro and simulate some of the biological characteristics of solid tumors, enhancing the evaluation of anticancer drug efficacy. Automated image analysis can be used for the assessment of tumor growth and documentation of changes in the size parameters of 3D tumor spheroids following anticancer treatments such as electrochemotherapy. The objective of this article is to assess the effect of various electroporation (EP) conditions (500–750 Volts/cm, 8–20 pulses, 100 µs pulse duration, 5 Hz repetition rate) combined with different bleomycin concentrations (1–2.5 ug/mL) on normal epithelial (HCjE-Gi) and conjunctival melanoma (CRMM1, CRMM2) 3D-cell cultures, through an automated image analysis and a comparison with standard histological assays. A reduction in tumor mass with loss of cell definition was observed after ECT (750 Volts/cm with eight pulses and 500 Volts/cm with 20 pulses) with bleomycin (1 μg/mL and 2.5 μg/mL) in the histological and immunohistochemical analyses of 3D CRMM1 and CRMM2 spheroids, whereas an increase in volume and a decrease in sphericity was documented in the automated image analysis and 3D visualization of both melanoma cell lines. For all other treatment conditions and for the HCjE-Gi cell line, no significant changes to their morphological features were observed. Image analysis with integrated software tools provides an accessible and comprehensive platform for the preliminary selection of homogenous spheroids and for the monitoring of drug efficacy, implementing the traditional screening methods. Full article
(This article belongs to the Special Issue 3D Cell and Tissue Culture)
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13 pages, 4496 KiB  
Article
Three-Dimensional Model of Dorsal Root Ganglion Explant as a Method of Studying Neurotrophic Factors in Regenerative Medicine
by Polina Klimovich, Kseniya Rubina, Veronika Sysoeva and Ekaterina Semina
Biomedicines 2020, 8(3), 49; https://doi.org/10.3390/biomedicines8030049 - 03 Mar 2020
Cited by 5 | Viewed by 5034
Abstract
Neurotrophic factors play a key role in the development, differentiation, and survival of neurons and nerve regeneration. In the present study, we evaluated the effect of certain neurotrophic factors (NGF, BDNF, and GDNF) on axon growth and migration of Nestin-green fluorescent protein (GFP)-positive [...] Read more.
Neurotrophic factors play a key role in the development, differentiation, and survival of neurons and nerve regeneration. In the present study, we evaluated the effect of certain neurotrophic factors (NGF, BDNF, and GDNF) on axon growth and migration of Nestin-green fluorescent protein (GFP)-positive cells using a 3D model of dorsal root ganglion (DRG) explant culture in Matrigel. Our method generally represents a convenient model for assessing the effects of soluble factors and therapeutic agents on axon growth and nerve regeneration in R&D studies. By analyzing the DRG explants in ex vivo culture for 21 days, one can evaluate the parameters of neurite outgrowth and the rate of cell migration from the DRG explants into the Matrigel. For the current study, we used Nestin-GFP-expressing mice in which neural precursors express Nestin and the green fluorescent protein (GFP) under the same promoter. We revealed that GDNF significantly (two fold) stimulated axon outgrowth (p < 0.05), but not BDNF or NGF. It is well-known that axon growth can be stimulated by activated glial cells that fulfill a trophic function for regenerating nerves. For this reason, we evaluated the number of Nestin-GFP-positive cells that migrated from the DRG into the Matrigel in our 3D ex vivo explant model. We found that NGF and GDNF, but not BDNF, stimulated the migration of Nestin-GFP cells compared to the control (p < 0.05). On the basis of the aforementioned finding, we concluded that GDNF had the greatest stimulating potential for axon regeneration, as it stimulated not only the axon outgrowth, but also glial cell migration. Although NGF significantly stimulated glial cell migration, its effect on axon growth was insufficient for axon regeneration. Full article
(This article belongs to the Special Issue 3D Cell and Tissue Culture)
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Review

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19 pages, 1420 KiB  
Review
3D Cell Culture Models in COVID-19 Times: A Review of 3D Technologies to Understand and Accelerate Therapeutic Drug Discovery
by Guadalupe Tonantzin de Dios-Figueroa, Janette del Rocío Aguilera-Marquez, Tanya A. Camacho-Villegas and Pavel H. Lugo-Fabres
Biomedicines 2021, 9(6), 602; https://doi.org/10.3390/biomedicines9060602 - 26 May 2021
Cited by 13 | Viewed by 10016
Abstract
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 [...] Read more.
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). Full article
(This article belongs to the Special Issue 3D Cell and Tissue Culture)
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18 pages, 3309 KiB  
Review
Utility of Three-Dimensional Cultures of Primary Human Hepatocytes (Spheroids) as Pharmacokinetic Models
by Kenta Mizoi, Hiroshi Arakawa, Kentaro Yano, Satoshi Koyama, Hajime Kojima and Takuo Ogihara
Biomedicines 2020, 8(10), 374; https://doi.org/10.3390/biomedicines8100374 - 23 Sep 2020
Cited by 17 | Viewed by 3530
Abstract
This paper reviews the usefulness, current status, and potential of primary human hepatocytes (PHHs) in three-dimensional (3D) cultures, also known as spheroids, in the field of pharmacokinetics (PK). Predicting PK and toxicity means pharmaceutical research can be conducted more efficiently. Various in vitro [...] Read more.
This paper reviews the usefulness, current status, and potential of primary human hepatocytes (PHHs) in three-dimensional (3D) cultures, also known as spheroids, in the field of pharmacokinetics (PK). Predicting PK and toxicity means pharmaceutical research can be conducted more efficiently. Various in vitro test systems using human hepatocytes have been proposed as tools to detect hepatic toxicity at an early stage in the drug development process. However, such evaluation requires long-term, low-level exposure to the test compound, and conventional screening systems such as PHHs in planar (2D) culture, in which the cells can only survive for a few days, are unsuitable for this purpose. In contrast, spheroids consisting of PHH are reported to retain the functional characteristics of human liver for at least 35 days. Here, we introduce a fundamental PK and toxicity assessment model of PHH spheroids and describe their applications for assessing species-specific metabolism, enzyme induction, and toxicity, focusing on our own work in these areas. The studies outlined in this paper may provide important information for pharmaceutical companies to reduce termination of development of drug candidates. Full article
(This article belongs to the Special Issue 3D Cell and Tissue Culture)
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