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Recent Advance in 3D Cultures
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Recent Advance in 3D Cultures

A special issue of International Journal of Molecular Sciences (ISSN 1422-0067). This special issue belongs to the section "Materials Science".

Deadline for manuscript submissions: closed (15 May 2023) | Viewed by 13344

Special Issue Editors

Dr. Isotta Chimenti

E-Mail Website
Guest Editor
Department of Medical and Surgical Sciences and Biotechnology, Faculty of Pharmacy and Medicine, Sapienza University of Rome, 04100 Latina, Italy
Interests: cardiac microenvironment; cardiac repair mechanisms; 3D culture
Special Issues, Collections and Topics in MDPI journals
Dr. Roberto Gaetani

E-Mail Website
Guest Editor
Department of Molecular Medicine, Università degli Studi di Roma La Sapienza, Rome, Italy
Interests: biomaterials; stem cells; cardiac regeneration; cardiac tissue engineering; tissue modelling; diabetes; microenvironment

Special Issue Information

Dear Colleagues,

Methods and protocols for creating 3D cultures in vitro have been rapidly evolving in recent years, concerning both biomaterials and structure design. Creating a 3D microenvironment for cell cultures allows complex interactions and stimuli that are functional for efficient phenotypic control and for mimicking tissue homeostasis and pathology. These tools can be used to successfully generate artificial tissues or cellular organoids that could be used for modelling the microenvironment in a physiologically relevant way, drug screening, or exploiting tissue engineering strategies in the clinical translation of regenerative medicine approaches.

This Special Issue will gather scholars in the field of 3D cultures for the creation of in vitro microenvironments for the study of tissue homeostasis and pathology, particularly in the field of cardiovascular research. Potential topics include, but are not limited to the following: novel methods for creating 3D cultures and tissue patches; study of the microenvironment in 3D cultures; modelling human diseases or pathological conditions in vitro; tissue engineering for regenerative medicine purposes; stem cells differentiation in 3D cultures; bioprinting of artificial tissues; development of smart biomaterials; cell–extracellular matrix interaction in the context of 3D tissue development and function.

This Special Issue is supervised by Dr. Isotta Chimenti, Dr. Roberto Gaetani, and assisted by Dr. Vittorio Picchio.

Dr. Isotta Chimenti
Dr. Roberto Gaetani
Guest Editors

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. International Journal of Molecular Sciences is an international peer-reviewed open access semimonthly journal published by MDPI.

Please visit the Instructions for Authors page before submitting a manuscript. There is an Article Processing Charge (APC) for publication in this open access journal. For details about the APC please see here. 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

  • 3D culture
  • tissue engineering
  • microenvironment
  • disease modelling
  • stem cells differentiation
  • regenerative medicine
  • biomaterials
  • cell-extracellular matrix interaction

Related Special Issue

Published Papers (7 papers)

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Editorial

Jump to: Research, Review

3 pages, 157 KiB  
Editorial
Recent Advances in 3D Cultures
by Vittorio Picchio, Roberto Gaetani and Isotta Chimenti
Int. J. Mol. Sci. 2024, 25(8), 4189; https://doi.org/10.3390/ijms25084189 - 10 Apr 2024
Viewed by 615
Abstract
Methods and protocols for creating complex 3D cell culture systems have been rapidly advancing in the past decade from the perspective of biomaterials [...] Full article
(This article belongs to the Special Issue Recent Advance in 3D Cultures)

Research

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21 pages, 15924 KiB  
Article
Elastomeric Porous Poly(glycerol sebacate) Methacrylate (PGSm) Microspheres as 3D Scaffolds for Chondrocyte Culture and Cartilage Tissue Engineering
by Dharaminder Singh, Sarah Lindsay, Shruti Gurbaxani, Aileen Crawford and Frederik Claeyssens
Int. J. Mol. Sci. 2023, 24(13), 10445; https://doi.org/10.3390/ijms241310445 - 21 Jun 2023
Cited by 2 | Viewed by 1412
Abstract
Cartilage defects can be difficult to treat; therefore, tissue engineering of cartilage is emerging as a promising potential therapy. One interesting area of research explores the delivery of cells to the cartilage defect via scaffold-based cell delivery vehicles and microsurgery. This study explores [...] Read more.
Cartilage defects can be difficult to treat; therefore, tissue engineering of cartilage is emerging as a promising potential therapy. One interesting area of research explores the delivery of cells to the cartilage defect via scaffold-based cell delivery vehicles and microsurgery. This study explores the use of novel poly(glycerol sebacate) methacrylate (PGSm)-polymerised high internal phase emulsion (polyHIPE) microspheres as scaffolds with embedded cells for cartilage tissue engineering. Porous microsphere scaffolds (100 µm–1 mm diameter) were produced from emulsions consisting of water and a methacrylate-based photocurable resin of poly(glycerol sebacate). These resins were used in conjunction with a T-junction fluidic device and an ultraviolet (UV) curing lamp to produce porous microspheres with a tuneable size. This technique produced biodegradable PGSm microspheres with similar mechanical properties to cartilage. We further explore these microspheres as scaffolds for three-dimensional culture of chondrocytes. The microspheres proved to be very efficient scaffolds for primary chondrocyte culture and were covered by a dense extracellular matrix (ECM) network during the culture period, creating a tissue disk. The presence of glycosaminoglycans (GAGs) and collagen-II was confirmed, highlighting the utility of the PGSm microspheres as a delivery vehicle for chondrocytes. A number of imaging techniques were utilised to analyse the tissue disk and develop methodologies to characterise the resultant tissue. This study highlights the utility of porous PGSm microspheres for cartilage tissue engineering. Full article
(This article belongs to the Special Issue Recent Advance in 3D Cultures)
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18 pages, 27695 KiB  
Article
3D ECM-Based Scaffolds Boost Young Cell Secretome-Derived EV Rejuvenating Effects in Senescent Cells
by Sharon Arcuri, Georgia Pennarossa, Teresina De Iorio, Fulvio Gandolfi and Tiziana A. L. Brevini
Int. J. Mol. Sci. 2023, 24(9), 8285; https://doi.org/10.3390/ijms24098285 - 5 May 2023
Cited by 3 | Viewed by 1755
Abstract
Aging is a complex, multifaceted degenerative process characterized by a progressive accumulation of macroscopic and microscopic modifications that cause a gradual decline of physiological functions. During the last few years, strategies to ease and counteract senescence or even rejuvenate cells and tissues were [...] Read more.
Aging is a complex, multifaceted degenerative process characterized by a progressive accumulation of macroscopic and microscopic modifications that cause a gradual decline of physiological functions. During the last few years, strategies to ease and counteract senescence or even rejuvenate cells and tissues were proposed. Here we investigate whether young cell secretome-derived extracellular vesicles (EVs) ameliorate the cellular and physiological hallmarks of aging in senescent cells. In addition, based on the assumption that extracellular matrix (ECM) provides biomechanical stimuli, directly influencing cell behavior, we examine whether ECM-based bio-scaffolds, obtained from decellularized ovaries of young swine, stably maintain the rejuvenated phenotype acquired by cells after exposure to young cell secretome. The results obtained demonstrate that young cells release EVs endowed with the ability to counteract aging. In addition, comparison between young and aged cell secretomes shows a significantly higher miR-200 content in EVs produced using fibroblasts isolated from young donors. The effect exerted by young cell secretome-derived EVs is transient, but can be stabilized using a young ECM microenvironment. This finding indicates a synergistic interaction occurring among molecular effectors and ECM-derived stimuli that cooperate to control a unique program, driving the cell clock. The model described in this paper may represent a useful tool to finely dissect the complex regulations and multiple biochemical and biomechanical cues driving cellular biological age. Full article
(This article belongs to the Special Issue Recent Advance in 3D Cultures)
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23 pages, 4290 KiB  
Article
Colorectal Cancer Bioengineered Microtissues as a Model to Replicate Tumor-ECM Crosstalk and Assess Drug Delivery Systems In Vitro
by Alessia La Rocca, Vincenza De Gregorio, Elena Lagreca, Raffaele Vecchione, Paolo Antonio Netti and Giorgia Imparato
Int. J. Mol. Sci. 2023, 24(6), 5678; https://doi.org/10.3390/ijms24065678 - 16 Mar 2023
Cited by 2 | Viewed by 1755
Abstract
Current 3D cancer models (in vitro) fail to reproduce complex cancer cell extracellular matrices (ECMs) and the interrelationships occurring (in vivo) in the tumor microenvironment (TME). Herein, we propose 3D in vitro colorectal cancer microtissues (3D CRC μTs), which reproduce the TME more [...] Read more.
Current 3D cancer models (in vitro) fail to reproduce complex cancer cell extracellular matrices (ECMs) and the interrelationships occurring (in vivo) in the tumor microenvironment (TME). Herein, we propose 3D in vitro colorectal cancer microtissues (3D CRC μTs), which reproduce the TME more faithfully in vitro. Normal human fibroblasts were seeded onto porous biodegradable gelatin microbeads (GPMs) and were continuously induced to synthesize and assemble their own ECMs (3D Stroma μTs) in a spinner flask bioreactor. Then, human colon cancer cells were dynamically seeded onto the 3D Stroma μTs to achieve the 3D CRC μTs. Morphological characterization of the 3D CRC μTs was performed to assess the presence of different complex macromolecular components that feature in vivo in the ECM. The results showed the 3D CRC μTs recapitulated the TME in terms of ECM remodeling, cell growth, and the activation of normal fibroblasts toward an activated phenotype. Then, the microtissues were assessed as a drug screening platform by evaluating the effect of 5-Fluorouracil (5-FU), curcumin-loaded nanoemulsions (CT-NE-Curc), and the combination of the two. When taken together, the results showed that our microtissues are promising in that they can help clarify complex cancer–ECM interactions and evaluate the efficacy of therapies. Moreover, they may be combined with tissue-on-chip technologies aimed at addressing further studies in cancer progression and drug discovery. Full article
(This article belongs to the Special Issue Recent Advance in 3D Cultures)
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Graphical abstract

14 pages, 6293 KiB  
Article
Optimization of Primary Human Bronchial Epithelial 3D Cell Culture with Donor-Matched Fibroblasts and Comparison of Two Different Culture Media
by Julian Maurer, Thorsten Walles and Cornelia Wiese-Rischke
Int. J. Mol. Sci. 2023, 24(4), 4113; https://doi.org/10.3390/ijms24044113 - 18 Feb 2023
Cited by 3 | Viewed by 1828
Abstract
In vitro airway models are increasingly important for pathomechanistic analyses of respiratory diseases. Existing models are limited in their validity by their incomplete cellular complexity. We therefore aimed to generate a more complex and meaningful three-dimensional (3D) airway model. Primary human bronchial epithelial [...] Read more.
In vitro airway models are increasingly important for pathomechanistic analyses of respiratory diseases. Existing models are limited in their validity by their incomplete cellular complexity. We therefore aimed to generate a more complex and meaningful three-dimensional (3D) airway model. Primary human bronchial epithelial cells (hbEC) were propagated in airway epithelial cell growth (AECG) or PneumaCult ExPlus medium. Generating 3D models, hbEC were airlifted and cultured on a collagen matrix with donor-matched bronchial fibroblasts for 21 days comparing two media (AECG or PneumaCult ALI (PC ALI)). 3D models were characterized by histology and immunofluorescence staining. The epithelial barrier function was quantified by transepithelial electrical resistance (TEER) measurements. The presence and function of ciliated epithelium were determined by Western blot and microscopy with high-speed camera. In 2D cultures, an increased number of cytokeratin 14-positive hbEC was present with AECG medium. In 3D models, AECG medium accounted for high proliferation, resulting in hypertrophic epithelium and fluctuating TEER values. Models cultured with PC ALI medium developed a functional ciliated epithelium with a stable epithelial barrier. Here, we established a 3D model with high in vivo–in vitro correlation, which has the potential to close the translational gap for investigations of the human respiratory epithelium in pharmacological, infectiological, and inflammatory research. Full article
(This article belongs to the Special Issue Recent Advance in 3D Cultures)
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15 pages, 1030 KiB  
Article
Beneficial Effect of Polysaccharide Gel Made of Xanthan Gum and Locust Bean Gum on Bovine Oocytes
by Shunsuke Hara, Yuki Inoue, Sogo Aoki, Keisuke Tanaka, Koumei Shirasuna and Hisataka Iwata
Int. J. Mol. Sci. 2023, 24(4), 3508; https://doi.org/10.3390/ijms24043508 - 9 Feb 2023
Cited by 4 | Viewed by 1490
Abstract
The present study examined the effect of polysaccharides gels made of xanthan gum and locust bean gum (gel culture system) on oocyte maturation and explored the molecules causing the beneficial effect of the gel culture system. Oocytes and cumulus cells complexes were collected [...] Read more.
The present study examined the effect of polysaccharides gels made of xanthan gum and locust bean gum (gel culture system) on oocyte maturation and explored the molecules causing the beneficial effect of the gel culture system. Oocytes and cumulus cells complexes were collected from slaughterhouse-derived ovaries and cultured on a plastic plate or gel. The gel culture system improved the rate of development to the blastocyst stage. The oocytes that matured on the gel contained high lipid contents and F-actin formation, and the resultant 8-cell stage embryos had low DNA methylation levels compared to their plate counterparts. RNA sequencing of the oocytes and embryos revealed the differentially expressed genes between the gel and plate culture systems, and upstream regulator analysis revealed estradiol and TGFB1 as top activated upstream molecules. The medium of the gel culture system contained higher concentrations of estradiol and TGFB1 than that of the plate cultures system. Supplementation of the maturation medium with either estradiol or TGFB1 resulted in high lipid content in oocytes. In addition, TGFB1 improved the developmental ability of the oocytes and increased F-actin content while reducing DNA methylation levels in the 8-cell stage embryos. In conclusion, the gel culture system is useful for embryo production, potentially through the upregulation of TGFB1. Full article
(This article belongs to the Special Issue Recent Advance in 3D Cultures)
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Review

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29 pages, 1924 KiB  
Review
Unlocking Neural Function with 3D In Vitro Models: A Technical Review of Self-Assembled, Guided, and Bioprinted Brain Organoids and Their Applications in the Study of Neurodevelopmental and Neurodegenerative Disorders
by Chiara D’Antoni, Lorenza Mautone, Caterina Sanchini, Lucrezia Tondo, Greta Grassmann, Gianluca Cidonio, Paola Bezzi, Federica Cordella and Silvia Di Angelantonio
Int. J. Mol. Sci. 2023, 24(13), 10762; https://doi.org/10.3390/ijms241310762 - 28 Jun 2023
Cited by 5 | Viewed by 3515
Abstract
Understanding the complexities of the human brain and its associated disorders poses a significant challenge in neuroscience. Traditional research methods have limitations in replicating its intricacies, necessitating the development of in vitro models that can simulate its structure and function. Three-dimensional in vitro [...] Read more.
Understanding the complexities of the human brain and its associated disorders poses a significant challenge in neuroscience. Traditional research methods have limitations in replicating its intricacies, necessitating the development of in vitro models that can simulate its structure and function. Three-dimensional in vitro models, including organoids, cerebral organoids, bioprinted brain models, and functionalized brain organoids, offer promising platforms for studying human brain development, physiology, and disease. These models accurately replicate key aspects of human brain anatomy, gene expression, and cellular behavior, enabling drug discovery and toxicology studies while providing insights into human-specific phenomena not easily studied in animal models. The use of human-induced pluripotent stem cells has revolutionized the generation of 3D brain structures, with various techniques developed to generate specific brain regions. These advancements facilitate the study of brain structure development and function, overcoming previous limitations due to the scarcity of human brain samples. This technical review provides an overview of current 3D in vitro models of the human cortex, their development, characterization, and limitations, and explores the state of the art and future directions in the field, with a specific focus on their applications in studying neurodevelopmental and neurodegenerative disorders. Full article
(This article belongs to the Special Issue Recent Advance in 3D Cultures)
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