Pre-validation of a Calu-3 epithelium cytotoxicity assay for predicting acute inhalation toxicity of chemicals

Jeong, Mi Ho; Han, Yan; Oh, In‐Sun; Kim, Dong-Min; Son, Dong–Won; Jung, Mi-Sook; Yang, Hyo-Seon; Lee, Kyuhong; Shin, Ju-Young; Kim, Ha Ryong; Chung, Kyu Hyuck

doi:10.1016/j.tiv.2021.105136

Cited by 1 publication

(1 citation statement)

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“…The use of primary lung cell cultures has not yet been well established, due to scarce availability of human tissue, different extraction techniques, donor‐to‐donor variability, short‐time viability, and complex cultivation methods of these cells (Silva et al, 2023). For lung epithelium assembly, the cell line derived from human bronchial adenocarcinoma (CALU‐3) has been widely used, since the behavior and phenotype of these cells fit the native tissue (Jeong et al, 2021; Kreft et al, 2015) in addition to resulting in high yields in culture (Pasman et al, 2021). Other cell lines such as BEAS‐2B and A549 can also be used in in vitro models.…”

Section: Introductionmentioning

confidence: 99%

CALU‐3 lung cells three‐dimensionally assembled onto CellFate® matrix present angiotensin‐converting enzyme‐2 activity

dos Santos,

dos Reis,

Berti

et al. 2023

Biotech & Bioengineering

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Currently, there is a great need for the development of three‐dimensional (3D) in vitro lung models. Particularly, the production of a suitable 3D model of pulmonary epithelium for understanding the pathophysiology of diseases such as the COVID‐19 must consider the tissue architecture and presence, for example, of the angiotensin‐converting enzyme‐2 (ACE‐2) in the cells. Different polymeric membranes are being used to support cell culturing, especially of lung cells, however, there is still no information about the culture of these cells onto bacterial nanocellulose (BNC) matrices. We have used the BNC matrix CellFate® as a support for the assembly of a 3D in vitro model of lung epithelium, composed of human lung fibroblasts (HLF) and lung adenocarcinoma cells (CALU‐3). CellFate® matrices were made from bacterial fermentation resulting in a natural and biocompatible biopolymer. Cells were cultured onto CellFate® and maintained in a 5% CO2 humidified atmosphere at 37°C. Cell viability was assessed by the resazurin method The samples were, then, exposed to the air–liquid interface (ALI), and histologically analyzed. ACE‐2 activity was verified on the hydrolyze of the fluorogenic substrate Mca‐APK(Dnp)‐OH, and its presence was evaluated by flow cytometry. The expression of the anionic transporter SLCO3A1 was evaluated by qPCR. Cell viability analysis indicates that CellFate® was not toxic to these cells. By flow cytometry, the presence of the ACE‐2 was identified in the CALU‐3 cells surface corroborating the results obtained from enzymatic activity analysis. The SLCO3A1 transporter expression was identified in cells cultured onto CellFate®, but not in cells cultured onto the transwell (control). CALU‐3 cells cultivated onto CellFate® resulted in a pseudostratified organization, a typical morphology of the human respiratory tract epithelium. The current model opens perspectives for studies involving physiological characterization, improving its relevance for the understanding of the pathophysiology of diseases as well as the response to drugs.

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