Three-dimensional (3D) cell culture systems have gained increasing interest in drug discovery and tissue engineering due to their evident advantages in providing more physiologically relevant information and more predictive data for in vivo tests. In this review, we discuss the characteristics of 3D cell culture systems in comparison to the two-dimensional (2D) monolayer culture, focusing on cell growth conditions, cell proliferation, population, and gene and protein expression profiles. The innovations and development in 3D culture systems for drug discovery over the past 5 years are also reviewed in the article, emphasizing the cellular response to different classes of anticancer drugs, focusing particularly on similarities and differences between 3D and 2D models across the field. The progression and advancement in the application of 3D cell cultures in cell-based biosensors is another focal point of this review.

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Three-Dimensional Cell Culture Systems and Their Applications in Drug Discovery and Cell-Based Biosensors

Many types of mammalian cells can aggregate and differentiate into 3-D multicellular spheroids when cultured in suspension or a nonadhesive environment. Compared to conventional monolayer cultures, multicellular spheroids resemble real tissues better in terms of structural and functional properties. Multicellular spheroids formed by transformed cells are widely used as avascular tumor models for metastasis and invasion research and for therapeutic screening. Many primary or progenitor cells on the other hand, show significantly enhanced viability and functional performance when grown as spheroids. Multicellular spheroids in this aspect are ideal building units for tissue reconstruction. Here we review the current understanding of multicellular spheroid formation mechanisms, their biomedical applications, and recent advances in spheroid culture, manipulation, and analysis techniques.

https://www.onlinelibrary.wiley.com/doi/pdf/10.1002/biot.1285

Recent advances in three-dimensional multicellular spheroid culture for biomedical research

http://csmres.co.uk/cs.public.upd/article-downloads/Three-dimensional-cell-culture.pdf

Three-dimensional cell culture: the missing link in drug discovery.

Opportunities and challenges for use of tumor spheroids as models to test drug delivery and efficacy.

Cell culture is a widely used in vitro tool for improving our understanding of cell biology, tissue morphology, and mechanisms of diseases, drug action, protein production and the development of tissue engineering. Most research regarding cancer biology is based on experiments using two-dimensional (2D) cell cultures in vitro. However, 2D cultures have many limitations, such as the disturbance of interactions between the cellular and extracellular environments, changes in cell morphology, polarity, and method of division. These disadvantages led to the creation of models which are more closely able to mimic conditions in vivo. One such method is three-dimensional culture (3D). Optimisation of the culture conditions may allow for a better understanding of cancer biology and facilitate the study of biomarkers and targeting therapies. In this review, we compare 2D and 3D cultures in vitro as well as different versions of 3D cultures.

https://www.archivesofmedicalscience.com/pdf-63906-57842?filename=2D and 3D cell cultures _.pdf

2D and 3D cell cultures - a comparison of different types of cancer cell cultures.

We describe the generation of microfluidic platforms for the co-culture of primary hepatocytes and endothelial cells; these platforms mimic the architecture of a liver sinusoid. This paper describes a progressional study of creating such a liver sinusoid on a chip system. Primary rat hepatocytes (PRHs) were co-cultured with primary or established endothelial cells in layers in single and dual microchannel configurations with or without continuous perfusion. Cell viability and maintenance of hepatocyte functions were monitored and compared for diverse experimental conditions. When primary rat hepatocytes were co-cultured with immortalized bovine aortic endothelial cells (BAECs) in a dual microchannel with continuous perfusion, hepatocytes maintained their normal morphology and continued to produce urea for at least 30 days. In order to demonstrate the utility of our microfluidic liver sinusoid platform, we also performed an analysis of viral replication for the hepatotropic hepatitis B virus (HBV). HBV replication, as measured by the presence of cell-secreted HBV DNA, was successfully detected. We believe that our liver model closely mimics the in vivo liver sinusoid and supports long-term primary liver cell culture. This liver model could be extended to diverse liver biology studies and liver-related disease research such as drug induced liver toxicology, cancer research, and analysis of pathological effects and replication strategies of various hepatotropic infectious agents. .

/pdf/liver-sinusoid-on-a-chip-long-term-layered-co-culture-of-icqj32iirk.pdf

Liver sinusoid on a chip: Long‐term layered co‐culture of primary rat hepatocytes and endothelial cells in microfluidic platforms

Micropatterns of Matrigel for three-dimensional epithelial cultures

Hepatocytes, the cells responsible for the metabolic and detoxification processes in the liver, are the predominant target of hepatitis B virus (HBV) infections, a major cause of liver cancer. The limited availability of normal human hepatocytes for cell-culture based studies is a significant challenge in HBV-associated liver cancer research. Therefore, there is a need for miniaturized cell-culture systems that can serve as a platform for studying the effect of HBV infections on hepatocyte physiology. Here, we present a microfluidic platform that can be used to study HBV replication in both rat and human hepatocytes. Polydimethylsiloxane (PDMS) microchannels fabricated using soft lithography techniques served as a culture vessel for both primary rat hepatocytes (PRH) and a human hepatoblastoma cell line, HepG2. The micro cell-culture chamber was then used as a model for HBV replication studies. Cells were grown in static culture conditions and either transfected with an HBV-genome cDNA or infected with the viral genome expressed from a recombinant adenovirus. Supernatants collected from the microchannels were assayed for secreted HBV using polymerase chain reaction (PCR). We achieved approximately 40 and 10% transfection efficiencies in HepG2 cells and PRH respectively, and 80–100% adenoviral infection efficiency in PRH comparable to standard tissue culture methods. Moreover, we successfully detected replicated HBV using our novel platform. This platform can be easily extended to studies involving DNA transfection or HBV infection of primary human hepatocytes since only a small number of cells are required for studies in microfluidic chambers.

Microfluidic platform for hepatitis B viral replication study

We have developed a novel microfluidic device using polydimethylsiloxane (PDMS) microchannels for long term, layered co-culture of primary rat hepatocytes (PRH) and endothelial cells (EC) to mimic the liver sinusoid. Three microfluidic configurations were investigated as shown in Figure 1. In configuration 1, where PRH and rat adrenal medullary endothelial cells (RAMEC) were co-cultured with matrigel in layers in a single PDMS microchannel, cells remained viable for 30 days. In configuration 3, where PRH and liver sinusoidal endothelial cells (LSEC) were cultured on the opposite sides of the microporous membrane between two microchannels under static and dynamic conditions, PRH retained their viability and normal morphology up to 35 days and 63 days, respectively. Thus, our novel liver models that closely mimic the liver sinusoid have been proven to facilitate long-term layered co-cultures of PRH and EC.

Temitope R Sodunke

Papers

Liver sinusoid on a chip: Long‐term layered co‐culture of primary rat hepatocytes and endothelial cells in microfluidic platforms

Micropatterns of Matrigel for three-dimensional epithelial cultures

Microfluidic platform for hepatitis B viral replication study

Liver on a chip: Engineering the liver sinusoid

Micropatterned matrigel track for 3D single breast cancer cell metastasis