ECVAM Good Cell Culture Practice Task Force Report 1

Hartung, Thomas; Balls, Michael; Bardouille, Claudia; Blanck, Olivier; Coecke, Sandra; Gstraunthaler, Gerhard; Lewis, David

doi:10.1177/026119290203000404

Cited by 89 publications

(47 citation statements)

References 20 publications

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“…Cell culture was carried out according to the principles of good cell culture practice (GCCP; Hartung et al 2002). Cells were maintained in culture in 75-cm 2 flasks, split 1:3 twice a week and used between passage 52 and 64.…”

Section: Cell Culturementioning

confidence: 99%

Comparison of the sensitivity of different toxicological endpoints in Caco-2 cells after cadmium chloride treatment

Boveri

Pazos

Gennari

et al. 2004

Archives of Toxicology

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The human colorectal adenocarcinoma cell line Caco-2 is a widely used in vitro model of the intestinal barrier. Cadmium chloride (CdCl(2)) is a highly toxic metal compound, ubiquitous in the biosphere, able to enter the food chain and to reach the intestinal epithelium, causing structural and functional damages. The aim of this work was to characterise cadmium toxicity in Caco-2 cells and, in particular, to compare the sensitivity of different endpoints revealing damage both on the epithelial barrier and at the cellular or molecular level. After 24-h exposure of the cells to CdCl(2), lactate dehydrogenase (LDH) leakage showed cadmium-induced cell toxicity, significant from 25 microM CdCl(2) and above, and analysis of different cell death pathways indicated the presence of necrosis after treatment with 50 microM CdCl(2). At the molecular level, we observed an increase in the protective protein heat shock protein 70 (HSP70), starting at 10 microM CdCl(2). At the barrier level, transepithelial electrical resistance (TEER) decreased while paracellular permeability (PCP) significantly increased after the treatment, showing an EC(50) of 6 and 16 microM CdCl(2), respectively, and indicating the loss of barrier integrity. In conclusion, our data reveal that CdCl(2) toxicity in Caco-2 cells can be detected at the barrier level at very low concentrations; also, HSP70 was shown to be a sensitive marker for detecting in vitro cadmium-induced toxicity.

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Section: Cell Culturementioning

confidence: 99%

Comparison of the sensitivity of different toxicological endpoints in Caco-2 cells after cadmium chloride treatment

Boveri

Pazos

Gennari

et al. 2004

Archives of Toxicology

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“…15 However, the decision as to when to terminate a certain stock of cells in culture and thaw another vial is often times made arbitrarily without any sound and solid scientific data. In our second example, we have shown that signature profiles can be used to assess HUVECs obtained from different suppliers, and in our hands they displayed significantly different profiles.…”

mentioning

confidence: 99%

Live Cell Quality Control and Utility of Real-Time Cell Electronic Sensing for Assay Development

Kirstein¹,

Atienza²,

Xi³

et al. 2006

ASSAY and Drug Development Technologies

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In this paper we have explored the utility of the real-time cell electronic sensing (RTCES, ACEA Biosciences Inc., San Diego, CA) system for monitoring the quality of live cells in cell-based assays as well as for assay development. We have demonstrated that each cell type displays unique growth kinetic profiles that provide a quantitative account of cell behavior and can be used as a diagnostic tool for cellular quality control. The utility of the specific signature patterns was shown by demonstrating the significant differences in primary cell behavior depending on the supplier. In addition, the RT-CES system was able to differentiate cell behavior depending on the passage stage of the cells. The utility of the RT-CES system as an assay development tool was demonstrated in cytotoxicity assays. The RT-CES system not only provides information regarding the potency of cytotoxic compounds, but in addition relates potency to the rate of the response for each concentration of the compound tested, which is important for understanding the mechanism of compound action. Moreover, real-time display of cytotoxicity data by the RT-CES system allows for calculation of real-time 50% inhibitory concentration (IC50) values or determination of optimal IC(50) value. In summary, the RT-CES system provides high content and information-rich data that are beyond the scope of single-point assays.

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“…It is important to highlight that to achieve reproducible results using our 3-D model the investigator must adhere to good cell culture guidelines 46–48 . It is crucial to systematically control cells for viability, mycoplasma contamination and changes in cell growth behavior.…”

Section: Discussionmentioning

confidence: 99%

Development of a Multicellular Three-dimensional Organotypic Model of the Human Intestinal Mucosa Grown Under Microgravity

Salerno-Gonçalves

Fasano

Sztein

2016

JoVE

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Because cells growing in a three-dimensional (3-D) environment have the potential to bridge many gaps of cell cultivation in 2-D environments (e.g., flasks or dishes). In fact, it is widely recognized that cells grown in flasks or dishes tend to de-differentiate and lose specialized features of the tissues from which they were derived. Currently, there are mainly two types of 3-D culture systems where the cells are seeded into scaffolds mimicking the native extracellular matrix (ECM): (a) static models and (b) models using bioreactors. The first breakthrough was the static 3-D models. 3-D models using bioreactors such as the rotating-wall-vessel (RWV) bioreactors are a more recent development. The original concept of the RWV bioreactors was developed at NASA’s Johnson Space Center in the early 1990s and is believed to overcome the limitations of static models such as the development of hypoxic, necrotic cores. The RWV bioreactors might circumvent this problem by providing fluid dynamics that allow the efficient diffusion of nutrients and oxygen. These bioreactors consist of a rotator base that serves to support and rotate two different formats of culture vessels that differ by their aeration source type: (1) Slow Turning Lateral Vessels (STLVs) with a co-axial oxygenator in the center, or (2) High Aspect Ratio Vessels (HARVs) with oxygenation via a flat, silicone rubber gas transfer membrane. These vessels allow efficient gas transfer while avoiding bubble formation and consequent turbulence. These conditions result in laminar flow and minimal shear force that models reduced gravity (microgravity) inside the culture vessel. Here we describe the development of a multicellular 3-D organotypic model of the human intestinal mucosa composed of an intestinal epithelial cell line and primary human lymphocytes, endothelial cells and fibroblasts cultured under microgravity provided by the RWV bioreactor.

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ECVAM Good Cell Culture Practice Task Force Report 1

Cited by 89 publications

References 20 publications

Comparison of the sensitivity of different toxicological endpoints in Caco-2 cells after cadmium chloride treatment

Comparison of the sensitivity of different toxicological endpoints in Caco-2 cells after cadmium chloride treatment

Live Cell Quality Control and Utility of Real-Time Cell Electronic Sensing for Assay Development

Development of a Multicellular Three-dimensional Organotypic Model of the Human Intestinal Mucosa Grown Under Microgravity

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