Gelatin-Methacryloyl (GelMA) Hydrogels with Defined Degree of Functionalization as a Versatile Toolkit for 3D Cell Culture and Extrusion Bioprinting

Gelatin-methacryloyl (GelMA) is a semi-synthetic hydrogel which consists of gelatin derivatized with methacrylamide and methacrylate groups. These hydrogels provide cells with an optimal biological environment (e.g., RGD motifs for adhesion) and can be quickly photo-crosslinked, which provides shape fidelity and stability at physiological temperature. In the present work, we demonstrated how GelMA hydrogels can be synthesized with a specific degree of functionalization (DoF) and adjusted to the intended application as a three-dimensional (3D) cell culture platform. The focus of this work lays on producing hydrogel scaffolds which provide a cell promoting microenvironment for human adipose tissue-derived mesenchymal stem cells (hAD-MSCs) and are conductive to their adhesion, spreading, and proliferation. The control of mechanical GelMA properties by variation of concentration, DoF, and ultraviolet (UV) polymerization conditions is described. Moreover, hAD-MSC cell viability and morphology in GelMA of different stiffness was evaluated and compared. Polymerized hydrogels with and without cells could be digested in order to release encapsulated cells without loss of viability. We also demonstrated how hydrogel viscosity can be increased by the use of biocompatible additives, in order to enable the extrusion bioprinting of these materials. Taken together, we demonstrated how GelMA hydrogels can be used as a versatile tool for 3D cell cultivation.

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Proliferation and Migration of Human Osteoblasts on Porous Three Dimensional Scaffolds

Materials Science Forum ◽

10.4028/www.scientific.net/msf.638-642.506 ◽

2010 ◽

Vol 638-642 ◽

pp. 506-511 ◽

Cited By ~ 5

Author(s):

Claudia Bergemann ◽

Ernst Dieter Klinkenberg ◽

Frank Lüthen ◽

Arne Weidmann ◽

Regina Lange ◽

...

Keyword(s):

Cell Culture ◽

Morphological Changes ◽

Three Dimensional ◽

3D Cell Culture ◽

Porous Tantalum ◽

Human Osteoblasts ◽

A Cell ◽

And Migration ◽

Migration Of Cells

Porous tantalum (Ta) biomaterial is designed to function as a scaffold for osseous ingrowths and has found applications in orthopedics. Integration of this Ta foam into the neighboring bone requires that osteoprogenitor cells attach to the implant, grow into the scaffold, proliferate and differentiate to osteoblasts. The aim of the present study was to create an in vitro 3D model system to investigate the interaction of human osteoblasts with porous Ta in the depth of the corpus. To explore active migration of osteoblasts into the Ta scaffold two porous Ta discs (Zimmer, Poland) were horizontally fixed within a clamping ring. Thereby a 3D Ta module with 4 levels is generated, which is placed into a cell culture well with the appropriate medium. Osteoblast-like cells were seeded apical onto the Ta module and cultured for 7 days in humidified atmosphere. Active migration of cells into the scaffold was monitored by field emission scanning electron microscopy (FESEM) imaging of the apical, medial and basal layers. A problem in 3D cell culture is the nutrition of cells inside of the scaffold. Therefore morphological changes and differentiation of the cells in distinct layers were analyzed.

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

International Journal of Molecular Sciences ◽

10.3390/ijms22052491 ◽

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.

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Mammary gland 3D cell culture systems in farm animals

Veterinary Research ◽

10.1186/s13567-021-00947-5 ◽

2021 ◽

Vol 52 (1) ◽

Author(s):

Laurence Finot ◽

Eric Chanat ◽

Frederic Dessauge

Keyword(s):

Cell Culture ◽

Mammary Gland ◽

Bacterial Infections ◽

Three Dimensional ◽

3D Cell Culture ◽

Farm Animals ◽

Culture Systems ◽

Cell Culture Systems

AbstractIn vivo study of tissue or organ biology in mammals is very complex and progress is slowed by poor accessibility of samples and ethical concerns. Fortunately, however, advances in stem cell identification and culture have made it possible to derive in vitro 3D “tissues” called organoids, these three-dimensional structures partly or fully mimicking the in vivo functioning of organs. The mammary gland produces milk, the source of nutrition for newborn mammals. Milk is synthesized and secreted by the differentiated polarized mammary epithelial cells of the gland. Reconstructing in vitro a mammary-like structure mimicking the functional tissue represents a major challenge in mammary gland biology, especially for farm animals for which specific agronomic questions arise. This would greatly facilitate the study of mammary gland development, milk secretion processes and pathological effects of viral or bacterial infections at the cellular level, all with the objective of improving milk production at the animal level. With this aim, various 3D cell culture models have been developed such as mammospheres and, more recently, efforts to develop organoids in vitro have been considerable. Researchers are now starting to draw inspiration from other fields, such as bioengineering, to generate organoids that would be more physiologically relevant. In this chapter, we will discuss 3D cell culture systems as organoids and their relevance for agronomic research.

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A rhabdomyosarcoma hydrogel model to unveil cell-extracellular matrix interactions

Biomaterials Science ◽

10.1039/d1bm00929j ◽

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)...

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Fabrication and characterization of egg white cryogel scaffold for three-dimensional (3D) cell culture

Biocatalysis and Agricultural Biotechnology ◽

10.1016/j.bcab.2018.12.019 ◽

2019 ◽

Vol 17 ◽

pp. 441-446 ◽

Cited By ~ 3

Author(s):

Perumalsamy Balaji ◽

Anbazhagan Murugadas ◽

Sellathamby Shanmugaapriya ◽

Mohammad Abdulkader Akbarsha

Keyword(s):

Cell Culture ◽

Three Dimensional ◽

3D Cell Culture ◽

Egg White ◽

Fabrication And Characterization

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Modulation of Matrix Softness and Interstitial Flow for 3D Cell Culture Using a Cell-Microenvironment-on-a-Chip System

ACS Biomaterials Science & Engineering ◽

10.1021/acsbiomaterials.6b00379 ◽

2016 ◽

Vol 2 (11) ◽

pp. 1968-1975 ◽

Cited By ~ 8

Author(s):

Nicholas Edwin Clay ◽

Kyeonggon Shin ◽

Altug Ozcelikkale ◽

Min Kyung Lee ◽

Max H. Rich ◽

...

Keyword(s):

Cell Culture ◽

3D Cell Culture ◽

Interstitial Flow ◽

Cell Microenvironment ◽

A Cell

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Hydrogel scaffolds composed of genetically synthesized self-assembling peptides for three-dimensional cell culture

Polymer Journal ◽

10.1038/pj.2012.216 ◽

2012 ◽

Vol 45 (5) ◽

pp. 504-508 ◽

Cited By ~ 6

Author(s):

Masayasu Mie ◽

Mayu Oomuro ◽

Eiry Kobatake

Keyword(s):

Cell Culture ◽

Three Dimensional ◽

Hydrogel Scaffolds ◽

Self Assembling ◽

Dimensional Cell

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Study on the Toxicity of Calix[4]- and [6]-Arenes and their Sulfated Derivatives in Two- and Three-Dimensional Cell Cultures of Colorectal Adenocarcinoma

Biotekhnologiya ◽

10.21519/0234-2758-2021-37-1-87-94 ◽

2021 ◽

Vol 37 (1) ◽

pp. 87-94

Author(s):

S.V. Nikulin ◽

B.Ya. Alekseev ◽

A.N Gorbunov ◽

I.M. Vatsuro ◽

V.V. Kovalev ◽

...

Keyword(s):

Cell Culture ◽

Colorectal Adenocarcinoma ◽

Three Dimensional ◽

3D Cell Culture ◽

Russian Science ◽

Adenocarcinoma Cells ◽

Therapeutic Molecules ◽

Derivatives Of ◽

Ht 29 ◽

Dimensional Cell

A comparative study of the toxicity of two unsubstituted calixarenes consisting of 4 and 6 phenolic fragments, as well as their p-sulfated derivatives, was carried out on the HT-29 colorectal adenocarcinoma cells cultured in two-dimensional (2D) and three-dimensional (3D) formats. It was shown that both unsubstituted calixarenes decrease the viability of tumor cells; calix[4]arene and calix[6]arene exhibited a cytostatic and a cytotoxic effect, respectively. Sulfated derivatives of calixarenes did not have a pronounced toxic effect on HT-29 cells. However, due to their high hydrophilicity and the ability to form adducts with various therapeutic molecules, they can be used for delivery of anticancer drugs. calixarenes, cytotoxicity, HT-29 cells, 2D cell culture, 3D cell culture The work was financially supported by the Russian Science Foundation (project no. 19-15-00397).

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The Late-Stage Protective Effect of Mito-TEMPO against Acetaminophen-Induced Hepatotoxicity in Mouse and Three-Dimensional Cell Culture Models

Antioxidants ◽

10.3390/antiox9100965 ◽

2020 ◽

Vol 9 (10) ◽

pp. 965

Author(s):

Mohammad Abdullah-Al-Shoeb ◽

Kenta Sasaki ◽

Saori Kikutani ◽

Nanami Namba ◽

Keiichi Ueno ◽

...

Keyword(s):

Oxidative Stress ◽

Cell Culture ◽

Liver Injury ◽

Protective Effect ◽

Acute Liver Injury ◽

Three Dimensional ◽

3D Cell Culture ◽

Cell Culture Model ◽

Culture Model ◽

Apap Hepatotoxicity

An overdose of acetaminophen (APAP), the most common cause of acute liver injury, induces oxidative stress that subsequently causes mitochondrial impairment and hepatic necroptosis. N-acetyl-L-cysteine (NAC), the only recognized drug against APAP hepatotoxicity, is less effective the later it is administered. This study evaluated the protective effect of mitochondria-specific Mito-TEMPO (Mito-T) on APAP-induced acute liver injury in C57BL/6J male mice, and a three dimensional (3D)-cell culture model containing the human hepatoblastoma cell line HepG2. The administration of Mito-T (20 mg/kg, i.p.) 1 h after APAP (400 mg/kg, i.p.) injection markedly attenuated the APAP-induced elevated serum transaminase activity and hepatic necrosis. However, Mito-T treatment did not affect key factors in the development of APAP liver injury including the activation of c-jun N-terminal kinases (JNK), and expression of the transcription factor C/EBP homologous protein (CHOP) in the liver. However, Mito-T significantly reduced the APAP-induced increase in the hepatic oxidative stress marker, nitrotyrosine, and DNA fragmentation. Mito-T markedly attenuated cytotoxicity induced by APAP in the HepG2 3D-cell culture model. Moreover, liver regeneration after APAP hepatotoxicity was not affected by Mito-T, demonstrated by no changes in proliferating cell nuclear antigen formation. Therefore, Mito-T was hepatoprotective at the late-stage of APAP overdose in mice.

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