An antioxidant self-healing hydrogel for 3D cell cultures

New insights into the biomechanical properties of cells are revealing the importance of these properties and how they relate to underlying molecular, architectural, and behavioral changes associated with cell state and disease processes. However, the current understanding of how these in vitro biomechanical properties are associated with in vivo processes has been developed based on the traditional monolayer (two-dimensional [2D]) cell culture, which traditionally has not translated well to the three-dimensional (3D) cell culture and in vivo function. Many gold standard methods and tools used to observe the biomechanical properties of 2D cell cultures cannot be used with 3D cell cultures. Fluorescent molecules can respond to external factors almost instantaneously and require relatively low-cost instrumentation. In this review, we provide the background on fluorescent molecular rotors, which are attractive tools due to the relationship of their emission quantum yield with environmental microviscosity. We make the case for their use in both 2D and 3D cell cultures and speculate on their fundamental and practical applications in cell biology.

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Upscaling of pneumatic membrane valves for the integration of 3D cell cultures on chip

Lab on a Chip ◽

10.1039/d1lc00194a ◽

2021 ◽

Author(s):

Nina Compera ◽

Scott Atwell ◽

Johannes Wirth ◽

Bernhard Wolfrum ◽

Matthias Meier

Keyword(s):

Cell Culture ◽

Cell Cultures ◽

Large Scale ◽

3D Cell Culture ◽

3D Cell Cultures ◽

Culture Formation ◽

Large Scale Integration ◽

3D Printed ◽

On Chip ◽

Scale Integration

For integration of 3D cell cultures on microfluidic large-scale integration chips, we upscaled pneumatic membrane valves using 3D-printed replica molds. Unit cell operations for 3D cell culture formation, culture, retrieval, and fusion are designed.

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Simplified One-stop Protocols to Extract Collagen from Various Animal Tissues for 3D Cell Cultures

10.26434/chemrxiv.13011536.v1 ◽

2020 ◽

Author(s):

John A. Terrell ◽

Chengpeng Chen

Keyword(s):

Cell Culture ◽

Cell Cultures ◽

3D Cell Culture ◽

Animal Tissues ◽

3D Cell Cultures ◽

One Stop

In this manuscript, we present a one-stop protocol to extract collagen from various animal tissues, and apply it to fabricate 3D cell culture scaffolds.

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Fluorination on polyethylenimine allows efficient 2D and 3D cell culture gene delivery

Journal of Materials Chemistry B ◽

10.1039/c4tb01447b ◽

2015 ◽

Vol 3 (4) ◽

pp. 642-650 ◽

Cited By ~ 34

Author(s):

Jia Lv ◽

Hong Chang ◽

Yu Wang ◽

Mingming Wang ◽

Jianru Xiao ◽

...

Keyword(s):

Cell Culture ◽

Gene Delivery ◽

Cell Cultures ◽

Gene Transfection ◽

3D Cell Culture ◽

Transfection Efficacy ◽

3D Cell Cultures ◽

2D And 3D

Fluorinationviaanhydride and oxirane reactions enhances the gene transfection efficacy of PEI on 3D cell cultures.

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Simplified One-stop Protocols to Extract Collagen from Various Animal Tissues for 3D Cell Cultures

10.26434/chemrxiv.13011536 ◽

2020 ◽

Author(s):

John A. Terrell ◽

Chengpeng Chen

Keyword(s):

Cell Culture ◽

Cell Cultures ◽

3D Cell Culture ◽

Animal Tissues ◽

3D Cell Cultures ◽

One Stop

In this manuscript, we present a one-stop protocol to extract collagen from various animal tissues, and apply it to fabricate 3D cell culture scaffolds.

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SpheroidPicker: An Automated 3D cell culture manipulator robot using deep learning

10.1101/2020.11.25.397331 ◽

2020 ◽

Author(s):

Istvan Grexa ◽

Akos Diosdi ◽

Andras Kriston ◽

Nikita Moshkov ◽

Maria Harmati ◽

...

Keyword(s):

Cell Culture ◽

Cell Cultures ◽

High Throughput Screening ◽

Ex Vivo ◽

Low Cost ◽

3D Cell Culture ◽

Experimental Conditions ◽

The Past ◽

3D Cell Cultures ◽

Manipulator Robot

AbstractRecent statistics report that more than 3.7 million new cases of cancer occur in Europe yearly, and the disease accounts for approximately 20 % of all deaths. High-throughput screening of cancer cell cultures has dominated the search for novel, effective anticancer therapies in the past decades. Recently, ex vivo 3D cell cultures from the patient’s own cancer cells have gained importance. We recently evaluated the major advancements and needs of the 3D cell cultures screening field, and we concluded that strictly standardized sample preparation is the most desired development. Here we propose an artificial intelligence-guided low-cost 3D cell culture delivery system. It consists of a light microscope, a micromanipulator, a syringe pump, and a controller computer. The system performs morphology-based feature analysis on spheroids and transfers the most appropriate ones between various sample holders. It can select the samples from standard sample holders, including Petri dishes and microwell plates, and then transfer them to a variety of holders up to 384 well plates. The device performs reliable semi- and fully automated spheroid transfer. This results in highly controlled experimental conditions and eliminates non-trivial side effects of sample variability that is a key aspect towards next-generation precision medicine.

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Three-Dimensional Cell Cultures as an In Vitro Tool for Prostate Cancer Modeling and Drug Discovery

International Journal of Molecular Sciences ◽

10.3390/ijms21186806 ◽

2020 ◽

Vol 21 (18) ◽

pp. 6806 ◽

Cited By ~ 2

Author(s):

Fabrizio Fontana ◽

Michela Raimondi ◽

Monica Marzagalli ◽

Michele Sommariva ◽

Nicoletta Gagliano ◽

...

Keyword(s):

Prostate Cancer ◽

Cell Culture ◽

Drug Discovery ◽

Cancer Cells ◽

Cell Cultures ◽

Antitumor Agents ◽

Three Dimensional ◽

3D Cell Culture ◽

Cancer Modeling

In the last decade, three-dimensional (3D) cell culture technology has gained a lot of interest due to its ability to better recapitulate the in vivo organization and microenvironment of in vitro cultured cancer cells. In particular, 3D tumor models have demonstrated several different characteristics compared with traditional two-dimensional (2D) cultures and have provided an interesting link between the latter and animal experiments. Indeed, 3D cell cultures represent a useful platform for the identification of the biological features of cancer cells as well as for the screening of novel antitumor agents. The present review is aimed at summarizing the most common 3D cell culture methods and applications, with a focus on prostate cancer modeling and drug discovery.

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Comparison of antioxidant activity between cyanidin-3-O-glucoside (C3G) liposome and cyanidin-3-O-glucoside (C3G) in 2D and 3D cell cultures

10.1101/314369 ◽

2018 ◽

Author(s):

Tisong Liang ◽

Rongfa Guan ◽

Guozhou Cao ◽

Haitao Shen ◽

Zhenfeng Liu ◽

...

Keyword(s):

Cell Culture ◽

Antioxidant Activity ◽

Cell Cultures ◽

Cell Model ◽

3D Cell Culture ◽

Culture Model ◽

Mda Content ◽

2D And 3D ◽

Cells Cultured

ABSTRACTThe 2D cell culture is the predominant in vitro model for numerous studies. However, 2D cell cultures may not accurately reflect the functions of three-dimensional (3D) tissues, which have extensive cell–cell and cell–matrix interactions; thus, using 2D cell cultures may lead to inaccurate experimental results. Therefore, to obtain adequate and detailed information about the antioxidant activity of cyanidin-3-O-glucoside (C3G) and C3G liposomes in the 2D and 3D cell culture models, we used in this study H2O2to construct the cell damage model and assess the antioxidant activity of C3G and C3G liposomes on Caco-2 cells cultured in the 3D model. We also measured the cell viability, cell morphology, and activity of glutathione (GSH), superoxide dismutase (SOD), total antioxidant capacity (T-AOC), and malondialdehyde (MDA) content of Caco-2 cells treated with H2O2, C3G, and C3G liposomes. Results showed that cells cultured in the 3D culture model formed a 3D structure and tight spheroids and showed increased cell activity and IC50. The C3G and C3G liposomes can enhance the activity of GSH, SOD, and T-AOC but decrease the MDA content. At the same time, the effect was more obvious in the 3D cell culture model than in the cells cultured in the 2D model. This study revealed that the results obtained from the 2D cell model may be inaccurate compared with the results obtained from the 3D cell model. A realistic mechanism study of antioxidant activity of C3G and C3G liposomes in the 3D cell model, which acts as an intermediate stage bridging the in vitro 2D and in vivo models, was observed.

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Gelatin-Methacryloyl (GelMA) Formulated with Human Platelet Lysate Supports Mesenchymal Stem Cell Proliferation and Differentiation and Enhances the Hydrogel’s Mechanical Properties

Bioengineering ◽

10.3390/bioengineering6030076 ◽

2019 ◽

Vol 6 (3) ◽

pp. 76 ◽

Cited By ~ 5

Author(s):

Marline Kirsch ◽

Luise Birnstein ◽

Iliyana Pepelanova ◽

Wiebke Handke ◽

Jessica Rach ◽

...

Keyword(s):

Cell Culture ◽

Stem Cell ◽

Mesenchymal Stem Cell ◽

Osteogenic Differentiation ◽

Cell Cultures ◽

Human Platelet ◽

3D Cell Culture ◽

Platelet Lysate ◽

Dependent Manner ◽

Human Platelet Lysate

Three-dimensional (3D) cell culture is a major focus of current research, since cultivation under physiological conditions provides more reliable information about in vivo cell behavior. 3D cell cultures are used in basic research to better understand intercellular and cell-matrix interactions. However, 3D cell culture plays an increasingly important role in the in vitro testing of bioactive substances and tissue engineering. Gelatin-methacryloyl (GelMA) hydrogels of different degrees of functionalization (DoFs) are a versatile tool for 3D cell culture and related applications such as bioprinting. Human platelet lysate (hPL) has already demonstrated positive effects on 2D cell cultures of different cell types and has proven a valuable alternative to fetal calf serum (FCS). Traditionally, all hydrogels are formulated using buffers. In this study, we supplemented GelMA hydrogels of different DoF with hPL during adipose tissue-derived mesenchymal stem cell (AD-MSCs) encapsulation. We studied the effect of hPL supplementation on the spreading, proliferation, and osteogenic differentiation of AD-MSCs. In addition, the influence of hPL on hydrogel properties was also investigated. We demonstrate that the addition of hPL enhanced AD-MSC spreading, proliferation, and osteogenic differentiation in a concentration-dependent manner. Moreover, the addition of hPL also increased GelMA viscosity and stiffness.

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Viscoelastic Oxidized Alginates with Reversible Imine Type Crosslinks: Self-Healing, Injectable, and Bioprintable Hydrogels

Gels ◽

10.3390/gels4040085 ◽

2018 ◽

Vol 4 (4) ◽

pp. 85 ◽

Cited By ~ 22

Author(s):

Shahzad Hafeez ◽

Huey Ooi ◽

Francis Morgan ◽

Carlos Mota ◽

Monica Dettin ◽

...

Keyword(s):

Mechanical Properties ◽

Cell Culture ◽

Viscoelastic Properties ◽

Soft Tissues ◽

3D Cell Culture ◽

Cell Delivery ◽

Self Healing ◽

Marked Influence ◽

Cell Culture Systems ◽

Multiscale Structures

Bioprinting techniques allow for the recreation of 3D tissue-like structures. By deposition of hydrogels combined with cells (bioinks) in a spatially controlled way, one can create complex and multiscale structures. Despite this promise, the ability to deposit customizable cell-laden structures for soft tissues is still limited. Traditionally, bioprinting relies on hydrogels comprised of covalent or mostly static crosslinks. Yet, soft tissues and the extracellular matrix (ECM) possess viscoelastic properties, which can be more appropriately mimicked with hydrogels containing reversible crosslinks. In this study, we have investigated aldehyde containing oxidized alginate (ox-alg), combined with different cross-linkers, to develop a small library of viscoelastic, self-healing, and bioprintable hydrogels. By using distinctly different imine-type dynamic covalent chemistries (DCvC), (oxime, semicarbazone, and hydrazone), rational tuning of rheological and mechanical properties was possible. While all materials showed biocompatibility, we observed that the nature of imine type crosslink had a marked influence on hydrogel stiffness, viscoelasticity, self-healing, cell morphology, and printability. The semicarbazone and hydrazone crosslinks were found to be viscoelastic, self-healing, and printable—without the need for additional Ca2+ crosslinking—while also promoting the adhesion and spreading of fibroblasts. In contrast, the oxime cross-linked gels were found to be mostly elastic and showed neither self-healing, suitable printability, nor fibroblast spreading. The semicarbazone and hydrazone gels hold great potential as dynamic 3D cell culture systems, for therapeutics and cell delivery, and a newer generation of smart bioinks.

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