Subacute cytotoxicity testing with cultured human lung cells

Yang, Aoran; Cardona, Delia; Barile, Frank A.

doi:10.1016/s0887-2333(01)00098-4

Cited by 16 publications

(10 citation statements)

References 6 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…As shown in Fig. 7(b), the cell inhibition rate had a similar trend compared to the result of a manual method and 99% confidence intervals (CI) (Yang et al 2002) of the IC 50 values were overlapped. For the result of the manual method, the IC 50 value was 1.6 µg/mL and the 99% CI was from 1.0 µg/mL to 2.8 µg/mL.…”

Section: Resultsmentioning

confidence: 54%

2-layer based microfluidic concentration generator by hybrid serial and volumetric dilutions

Lee

Kim

et al. 2010

Biomed Microdevices

View full text Add to dashboard Cite

We present a 2-layer based microfluidic concentration generator by a hybrid of a serial and a volumetric dilution for dose-response experiments in drug screening. The hybrid dilution method using 2-layer based microfluidic network significantly reduces the total number of cascaded serial dilution stages. The proposed strategy is capable of generating a large number of universal stepwise monotonic concentrations with a wide range of logarithmic and linear scales. We have studied an equivalent electrical circuit to that of the 2-layer based microfluidic network, where the only variable parameter is channel length. We have designed a microfluidic dilution generator simultaneously covering 14 doses with a combination of 4-order logarithmic and 4-point linear concentrations. The design has been verified by a commercial circuit analysis software (e.g., P-Spice) for the electrical circuit analysis and a computational fluid dynamics software (e.g., CFD-ACE+) for the microfluidic circuit analysis. As a real-life application of the proposed dilution generator, we have successfully performed a dose-response experiment using MCF-7 human breast cancer cells. We expect that the proposed dilution method will be useful to study not only high throughput drug screening but also optimization in biology, chemistry, medicine, and material sciences.

show abstract

Section: Resultsmentioning

confidence: 54%

2-layer based microfluidic concentration generator by hybrid serial and volumetric dilutions

Lee

Kim

et al. 2010

Biomed Microdevices

View full text Add to dashboard Cite

show abstract

“…As noted above, ethanol did not show toxicity as the negative control chemical at 100 mmol/L (0.5% w/v). We have also previously reported the IC 50 for ethanol in a 24-hr MTT assay using human lung cells at greater than 500 mmol/L (Yang et al, 2002). Table 2 summarizes the statistical analysis of the data in Table 1.…”

Section: Cytotoxicity Testing With Confluent Mes Cells: Comparison Ofmentioning

confidence: 99%

Evaluation of in vitro cytotoxicity and paracellular permeability of intact monolayers with mouse embryonic stem cells

Calabro

Konsoula

Barile

2008

Toxicology in Vitro

View full text Add to dashboard Cite

Mouse embryonic stem (mES) cells were induced to form intact monolayers in cell culture inserts, using combinations of extracellular matrix (ECM) components and growth factors (GFs). Progressive formation of intact monolayers was monitored using transepithelial electrical resistance (TEER) and passage of paracellular permeability (PP) markers. The mES cells were initially inoculated on inactivated mouse embryonic fibroblasts (MEFs) plus leukemia inhibitory factor (LIF). At 75% confluence, cells were passaged in the absence of MEF and LIF to stimulate formation of rounded multicellular aggregates (MA). After 4 days, cultures containing MA were transferred to culture inserts coated with ECM components only, and grown in the presence of selected individual GFs. An additional 10 to 14 days revealed confluent monolayers with TEER values of 500-700 ohmscm 2 (Ω-cm 2 ). Monolayers grown on inserts coated with ECM components, such as fibronectin or collagen-IV, in the presence of epidermal growth factor or keratinocyte growth factor in the medium, yielded the highest TEER measurements when compared to cultures grown without GFs or ECM. Acute cytotoxicity (AC) studies with confluent monolayers of mES cells in 96-well plates indicated that there is a high correlation (R 2 =0.91) between cell viability and TEER for 24-h exposure time. Also, decrease in TEER is inversely proportional with increase in PP of markers. In comparison to standardized Registry of Cytotoxicity (RC) data and TEER measurements, MTT IC 50 values for mES cells are lower. Thus, at equivalent concentrations for the same chemicals, cell viability decreases before the integrity of the monolayer is compromised. This system represents a novel approach for the manipulation of mES cells toward specific intact monolayers, as an in vitro model for biological monolayer formation, and most importantly, for applications to cytotoxicity testing.

show abstract

“…In vitro cytotoxicity models are often used to screen for potential harmful effects of chemical compounds because they have good reproducibility and sensitivity, while also minimizing the use of animals. Moreover, cellular cytotoxicity systems are better predictive tools for human toxicity than are whole organism models [5,6,15,16].…”

Section: Discussionmentioning

confidence: 99%

In VitroEvaluation of a Biomedical-Grade Bilayer Chitosan Porous Skin Regenerating Template as a Potential Dermal Scaffold in Skin Tissue Engineering

Lim

Halim

Zainol

et al. 2011

International Journal of Polymer Science

View full text Add to dashboard Cite

Chitosan is a copolymer of N-acetylglucosamine and glucosamine. A bilayer chitosan porous skin regenerating template (CPSRT) has been developed for skin tissue engineering. The pore size of the CPSRT was assessed using a scanning electron microscopy (SEM). The in vitro cytocompatibility of the CPSRT was tested on primary human epidermal keratinocyte (pHEK) cultures by measuring lactate dehydrogenase (LDH) levels and skin irritation by western blot analysis of the interleukin-8 (IL-8) and tumor necrosis factor-α (TNF-α) secretions. The ability of the CPSRT to support cell ingrowth was evaluated by seeding primary human dermal fibroblasts (pHDFs) on the scaffold, staining the cells with live/dead stain, and imaging the construct by confocal microscopy (CLSM). The CPSRT with pore sizes ranging from 50 to 150 μm was cytocompatible because it did not provoke the additional production of IL-8 and TNF-α by pHEK cultures. Cultured pHDFs were able to penetrate the CPSRT and had increased in number on day 14. In conclusion, the CPSRT serves as an ideal template for skin tissue engineering.

show abstract

Subacute cytotoxicity testing with cultured human lung cells

Cited by 16 publications

References 6 publications

2-layer based microfluidic concentration generator by hybrid serial and volumetric dilutions

2-layer based microfluidic concentration generator by hybrid serial and volumetric dilutions

Evaluation of in vitro cytotoxicity and paracellular permeability of intact monolayers with mouse embryonic stem cells

In VitroEvaluation of a Biomedical-Grade Bilayer Chitosan Porous Skin Regenerating Template as a Potential Dermal Scaffold in Skin Tissue Engineering

Contact Info

Product

Resources

About