Hanging drop: An in vitro air toxic exposure model using human lung cells in 2D and 3D structures

Liu, Fei; Peng, Cheng; Escher, Beate I.; Fantino, Emmanuelle; Giles, Cindy; Were, Stephen T.; Duffy, Lesley L.; Ng, J. C.

doi:10.1016/j.jhazmat.2013.01.027

Cited by 32 publications

(45 citation statements)

References 42 publications

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“…P.S. Hiemstra et al Toxicology in Vitro 47 (2018) 137-146 allowing direct contact between benzene in the air and the cells (Liu et al, 2013).…”

Section: Organoidsmentioning

confidence: 99%

“…However, duration of exposure beyond 6 h is a limitation in ALI systems, partly because of drying of the epithelial surface resulting in cellular stress and cell death. As an alternative, hanging drop (HD) culturing was used to produce spheroids from A549 cells, followed by exposure using sealed conditions to allow long-term (up to 20 days) exposures to even volatile compounds (Liu et al, 2013). The results of the exposure were more variable with A549 cultures precultured before exposure for 7 days (designated 3D by the authors) than after 1 day (designated 2D).…”

Section: Culture Models Of Lung Epithelial Cells: Exposure Systemsmentioning

confidence: 99%

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Human lung epithelial cell cultures for analysis of inhaled toxicants: Lessons learned and future directions

Hiemstra

Grootaers

Does

et al. 2018

Toxicology in Vitro

153

116

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A B S T R A C TThe epithelium that covers the conducting airways and alveoli is a primary target for inhaled toxic substances, and therefore a focus in inhalation toxicology. The increasing concern about the use of animal models has stimulated the development of in vitro cell culture models for analysis of the biological effects of inhaled toxicants. However, the validity of the current in vitro models and their acceptance by regulatory authorities as an alternative to animal models is a reason for concern, and requires a critical review. In this review, focused on human lung epithelial cell cultures as a model for inhalation toxicology, we discuss the choice of cells for these models, the cell culture system used, the method of exposure as well as the various read-outs to assess the cellular response. We argue that rapid developments in the 3D culture of primary epithelial cells, the use of induced pluripotent stem cells for generation of lung epithelial cells and the development of organ-on-a-chip technology are among the important developments that will allow significant advances in this field. Furthermore, we discuss the various routes of application of inhaled toxicants by air-liquid interface models as well as the vast array of read-outs that may provide essential information. We conclude that close collaboration between researchers from various disciplines is essential for development of valid methods that are suitable for replacement of animal studies for inhalation toxicology.

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“…P.S. Hiemstra et al Toxicology in Vitro 47 (2018) 137-146 allowing direct contact between benzene in the air and the cells (Liu et al, 2013).…”

Section: Organoidsmentioning

confidence: 99%

Section: Culture Models Of Lung Epithelial Cells: Exposure Systemsmentioning

confidence: 99%

Human lung epithelial cell cultures for analysis of inhaled toxicants: Lessons learned and future directions

Hiemstra

Grootaers

Does

et al. 2018

Toxicology in Vitro

153

116

View full text Add to dashboard Cite

show abstract

“…Moreover, after the culture, hydrogel removal is required to harvest the tumor spheroids for subsequent gene and protein assays. Thus, other 3D culture techniques have been developed to simplify the experimental processing, such as the liquid overlay technique, hanging drop method, and microfluidic‐based methods . Among these, the liquid overlay technique was adopted in the current study because the process is simple and compatible with the conventional biological skillset.…”

Section: Introductionmentioning

confidence: 99%

Apoptosis and cell cycle arrest of hepatocellular carcinoma spheroids treated by an alternating electric field

Huang

Lei

Tsang

2019

Biotechnology Progress

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Most of the current cancer therapies may induce serious side effects and affect patient quality of life. Recently, a novel treatment using an alternating low-intensity and intermediate-frequency electric field was proposed and found to be a noninvasive and minimally toxic approach. However, additional fundamental studies and scientific evidence are required to further support the development of this treatment into a standard cancer therapy. In the current work, an in-house fabricated culture plate was developed to study the responses of hepatocellular carcinoma spheroids to treatment with an alternating electric field. From the results of the viability study, the electric field was confirmed to influence the dividing cells in the spheroids. Fluorescent staining of live and dead cells revealed that a fraction of the cells were damaged in the fieldtreated spheroids. Moreover, flow cytometry analyses were conducted and showed that a fraction of the cells in the spheroids underwent apoptosis and cell cycle arrest.Additionally, the apoptosis pathway (Bax/caspase) and cell cycle arrest pathway (p53/p21) were found to be activated after exposure to the electric field. In summary, the results further elucidated the cellular and molecular mechanism inducing apoptosis and cell cycle arrest in the field-treated hepatocellular carcinoma spheroids. This study provides more evidence to support the efficacy of electric-field-based cancer therapy. K E Y W O R D Sapoptosis, cancer therapy, cell cycle arrest, electrical field, hepatocellular carcinoma spheroids

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“…Additionally, changing the composition of the culture medium or adding more cells to the aggregates are not feasible with this method. Moreover, this method has a narrow range of islet cell aggregate size and has not shown the flexibility to create different sizes (Banerjee & Bhonde, 2006;Cavallari et al, 2007;Liu et al, 2013;Pethig et al, 2008). These challenges could be addressed by combining the hanging-drop approach with the emerging cell printing technique.…”

Section: Hanging-drop Methodsmentioning

confidence: 99%

Engineering of microscale three-dimensional pancreatic islet modelsin vitroand their biomedical applications

Gao

Wang

Han

et al. 2015

Critical Reviews in Biotechnology

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Diabetes now is the most common chronic disease in the world inducing heavy burden for the people's health. Based on this, diabetes research such as islet function has become a hot topic in medical institutes of the world. Today, in medical institutes, the conventional experiment platform in vitro is monolayer cell culture. However, with the development of micro- and nano-technologies, several microengineering methods have been developed to fabricate three-dimensional (3D) islet models in vitro which can better mimic the islet of pancreases in vivo. These in vitro islet models have shown better cell function than monolayer cells, indicating their great potential as better experimental platforms to elucidate islet behaviors under both physiological and pathological conditions, such as the molecular mechanisms of diabetes and clinical islet transplantation. In this review, we present the state-of-the-art advances in the microengineering methods for fabricating microscale islet models in vitro. We hope this will help researchers to better understand the progress in the engineering 3D islet models and their biomedical applications such as drug screening and islet transplantation.

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Hanging drop: An in vitro air toxic exposure model using human lung cells in 2D and 3D structures

Cited by 32 publications

References 42 publications

Human lung epithelial cell cultures for analysis of inhaled toxicants: Lessons learned and future directions

Human lung epithelial cell cultures for analysis of inhaled toxicants: Lessons learned and future directions

Apoptosis and cell cycle arrest of hepatocellular carcinoma spheroids treated by an alternating electric field

Engineering of microscale three-dimensional pancreatic islet modelsin vitroand their biomedical applications

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