A Robust Protocol for Decellularized Human Lung Bioink Generation Amenable to 2D and 3D Lung Cell Culture

Decellularization efforts must balance the preservation of the extracellular matrix (ECM) components while eliminating the nucleic acid and cellular components. Following effective removal of nucleic acid and cell components, decellularized ECM (dECM) can be solubilized in an acidic environment with the assistance of various enzymes to develop biological scaffolds in different forms, such as sheets, tubular constructs, or three-dimensional (3D) hydrogels. Each organ or tissue that undergoes decellularization requires a distinct and optimized protocol to ensure that nucleic acids are removed, and the ECM components are preserved. The objective of this study was to optimize the decellularization process for dECM isolation from human lung tissues for downstream 2D and 3D cell culture systems. Following protocol optimization and dECM isolation, we performed experiments with a wide range of dECM concentrations to form human lung dECM hydrogels that were physically stable and biologically responsive. The dECM based-hydrogels supported the growth and proliferation of primary human lung fibroblast cells in 3D cultures. The dECM is also amenable to the coating of polyester membranes in Transwell™ Inserts to improve the cell adhesion, proliferation, and barrier function of primary human bronchial epithelial cells in 2D. In conclusion, we present a robust protocol for human lung decellularization, generation of dECM substrate material, and creation of hydrogels that support primary lung cell viability in 2D and 3D culture systems

Download Full-text

Practical Applications of 3D Cell Culture Biotechnologies for Translational Oncology and Personalized Therapy

Biotekhnologiya ◽

10.21519/0234-2758-2020-36-3-3-15 ◽

2020 ◽

Vol 36 (3) ◽

pp. 3-15

Author(s):

D.A. Chudakova ◽

E.Yu. Skorova ◽

I.V. Reshetov

Keyword(s):

Cell Culture ◽

Extracellular Matrix ◽

3D Culture ◽

3D Cell Culture ◽

Personalized Therapy ◽

3D Bioprinting ◽

Culture Systems ◽

Culture Models ◽

2D And 3D

The creation of in vitro three-dimensional cellular model systems (in vitro 3D cultures) is a fast-growing leading-edge segment of the biotechnological industry. We have examined in this work the key 80 articles published after 2008, and focused on applications of in vitro 3D culture in translational oncology. We described a broad range of 3D culture systems, including models with and without extracellular matrix (ECM). 3D culture models based on decellularized ECM were discussed in more detail. The role of ECM in pathogeneis of malignant neoplasms, in particular, in the phenomenon of the tumor resistance to chemotherapy, was evaluated. 2D and 3D culture systems were compared, and natural and synthetic ECM were described, as well as the model creation based on 3D bioprinting. Particular attention was paid to in vitro models of various cancers, including those at the metastatic stage, based on 3D cell cultures, which maximally mimic the in vivo tumor behavior. The prospects of the practical application of 3D cell culture models in preclinical drag screening and in personalized therapy were discussed. We also presented our data on in vitro 2D and 3D culturing of human cells on various substrates. 3D cellular models, 3D bioprinting, biotechnology, extracellular matrix, cancer, translational medicine, personalized medicine, drag development, in vitro, ex vivo, oncology The authors are grateful to Dr. E. Shabalina for providing part of the experimental data and to OKA-Biotech Company for the samples of recombinant Funding-The work was supported by a Grant from the Russian Science Foundation (no. 18-15-00391). doi: 10.21519/0234-2758-2020-36-3-3-15

Download Full-text

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)...

Download Full-text

Antitumor activity of amidino-substituted benzimidazole and benzimidazo[1,2-a]quinoline derivatives tested in 2D and 3D cell culture systems

Journal of Enzyme Inhibition and Medicinal Chemistry ◽

10.3109/14756366.2015.1101093 ◽

2015 ◽

Vol 31 (6) ◽

pp. 1139-1145 ◽

Cited By ~ 8

Author(s):

Karmen Brajša ◽

Ines Vujasinović ◽

Dubravko Jelić ◽

Marija Trzun ◽

Ivo Zlatar ◽

...

Keyword(s):

Cell Culture ◽

Antitumor Activity ◽

3D Cell Culture ◽

Quinoline Derivatives ◽

Culture Systems ◽

Cell Culture Systems ◽

2D And 3D

Download Full-text

Label-free and real-time monitoring of human mesenchymal stem cell differentiation in 2D and 3D cell culture systems using impedance cell sensors

RSC Advances ◽

10.1039/c8ra05273e ◽

2018 ◽

Vol 8 (54) ◽

pp. 31246-31254 ◽

Cited By ~ 7

Author(s):

Jun Ho Song ◽

Sun-Mi Lee ◽

Kyung-Hwa Yoo

Keyword(s):

Cell Culture ◽

Real Time ◽

3D Cell Culture ◽

Stem Cell Differentiation ◽

Human Mesenchymal Stem Cell ◽

Label Free ◽

Culture Systems ◽

Mesenchymal Stem Cell Differentiation ◽

Cell Culture Systems ◽

2D And 3D

3D impedance cell sensors are developed to monitor hMSC differentiation in label-free and real-time. Analyzing capacitance and conductance with these sensors shows that osteoblast and adipocyte lineages can be discriminated non-invasively in 3D cell culture systems.

Download Full-text

Alginate-based 3D cell culture technique to evaluate the half-maximal inhibitory concentration: an in vitro model of anticancer drug study for anaplastic thyroid carcinoma

Thyroid Research ◽

10.1186/s13044-021-00118-w ◽

2021 ◽

Vol 14 (1) ◽

Author(s):

Hilda Samimi ◽

Alireza Naderi Sohi ◽

Shiva Irani ◽

Ehsan Arefian ◽

Mojdeh Mahdiannasser ◽

...

Keyword(s):

Cell Culture ◽

Thyroid Carcinoma ◽

Cell Lines ◽

Cytotoxic Effect ◽

Inhibitory Concentration ◽

3D Culture ◽

3D Cell Culture ◽

Anaplastic Thyroid Carcinoma ◽

2D And 3D

Abstract Background Three-dimensional (3D) cell culture methods are identified for simulating the biological microenvironment and demonstrating more similarity to in vivo circumstances. Anaplastic thyroid carcinoma (ATC) is a lethal endocrine malignancy. Despite different treatment approaches, no improvement in the survival rate of the patients has been shown. In this study, we used the 3D in vitro ATC model to investigate the cytotoxic effect of BI-847325 anticancer drug in two-dimensional (2D)- and 3D- cultured cells. Methods Human ATC cell lines, C643 and SW1736, were cultured in one percentage (w/v) sodium alginate. Spheroids were incubated in medium for one week. The reproducibility of the fabrication of alginate beads was evaluated. Encapsulation of the cells in alginate was examined by DAPI (4′,6-diamidino-2-phenylindole) staining. Survival of alginate-encapsulated cells was evaluated by CFSE (5,6-Carboxyfluorescein N-hydroxysuccinimidyl ester) staining. The population doubling times of C643 and SW1736 cell lines cultured in 2D monolayer as well as in 3D system were calculated. The cytotoxic effect of BI-847325 on 2D- and 3D- cultured cell lines was assessed for 24–72 h by MTT [3-(4,5-dimethylthiazolyl-2)-2,5-diphenyltetrazolium bromide] assay. Finally, the 3D culture results were compared with the 2D culture method. Results The half-maximal inhibitory concentration (IC50) values of BI-847325 were higher in 3D culture compared to 2D culture. The cytotoxicity data indicated that 3D in vitro models were more resistant to chemotherapy agents. Conclusions The findings of this study are beneficial for developing in vitro ATC 3D models to analyze the efficacy of different chemotherapy drugs and formulations.

Download Full-text

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.

Download Full-text

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.

Download Full-text