Skip Navigation


ToxSci Advance Access originally published online on December 7, 2005
Toxicological Sciences 2006 90(1):133-141; doi:10.1093/toxsci/kfj067
This Article
Right arrow Full Text Freely available
Right arrow FREE Full Text (PDF) Freely available
Right arrow All Versions of this Article:
90/1/133    most recent
kfj067v1
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in ISI Web of Science
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Add to My Personal Archive
Right arrow Download to citation manager
Right arrow Search for citing articles in:
ISI Web of Science (11)
Right arrowRequest Permissions
Right arrow Disclaimer
Google Scholar
Right arrow Articles by Kasahara, T.
Right arrow Articles by Kakinuma, C.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Kasahara, T.
Right arrow Articles by Kakinuma, C.
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us  
What's this?

© The Author 2005. Published by Oxford University Press on behalf of the Society of Toxicology. All rights reserved. For Permissions, please email: journals.permissions@oxfordjournals.org

Establishment of an In Vitro High-Throughput Screening Assay for Detecting Phospholipidosis-Inducing Potential

Toshihiko Kasahara*,1, Kazuo Tomita*, Hiroyuki Murano{dagger}, Tsuyoshi Harada{ddagger}, Keisuke Tsubakimoto*, Takuo Ogihara*, Syuhei Ohnishi* and Chihaya Kakinuma*

* Pharmaceutical Research Center, {dagger} Research Planning and Management, and {ddagger} Development Research, Mochida Pharmaceutical Company Limited, Shizuoka, Japan

Received August 29, 2005; accepted November 16, 2005

Excessive accumulation of phospholipids results in phospholipidosis (PL), which may interfere with cellular functions, leading to acute or chronic disease or even death. Electron-microscopic detection of cytoplasmic lamellar bodies is often used as a diagnostic criterion of PL, but a faster, more convenient procedure is required for high-throughput assay of the PL-inducing potential of candidate drugs. We have developed a 96-well microplate cell-culture method for detecting PL, using a phosphatidylcholine-conjugated dye (NBD-PC) and a fluoro-microplate reader. The fluorescence intensity due to NBD-PC was normalized to that of Hoechst33342, used as an indicator of cell number, to obtain the amount of NBD-PC taken up per living cell. To select a suitable cell type, we examined the PL-detection sensitivity of five cell lines, as well as human and rat primary hepatocyte cultures, with five cationic amphiphilic drugs (CAD) as PL inducers and a negative control compound. The cell lines CHO-K1 and CHL/IU gave the best results. The NBD-PC uptake per CHO-K1 cell showed a high correlation with the pathological score of PL for 24 compounds, including PL-positive and negative compounds. This high-throughput screening assay for PL-inducing potential (HTS-PL assay) offers high sensitivity and accuracy, and it allows simultaneous determination of cytotoxicity.

Key Words: phospholipidosis; cationic amphiphilic drug; NBD-PC; Hoechst33342; HTS-PL assay.


Add to CiteULike CiteULike   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us    What's this?


This article has been cited by other articles:


Home page
 Toxicol SciHome page
P. Nioi, B. K. Perry, E.-J. Wang, Y.-Z. Gu, and R. D. Snyder
In Vitro Detection of Drug-Induced Phospholipidosis Using Gene Expression and Fluorescent Phospholipid Based Methodologies
Toxicol. Sci., September 1, 2007; 99(1): 162 - 173.
[Abstract] [Full Text] [PDF]

Home page
 Toxicol SciHome page
F. Atienzar, H. Gerets, S. Dufrane, K. Tilmant, M. Cornet, S. Dhalluin, B. Ruty, G. Rose, and M. Canning
Determination of Phospholipidosis Potential Based on Gene Expression Analysis in HepG2 Cells
Toxicol. Sci., March 1, 2007; 96(1): 101 - 114.
[Abstract] [Full Text] [PDF]


Disclaimer:
Please note that abstracts for content published before 1996 were created through digital scanning and may therefore not exactly replicate the text of the original print issues. All efforts have been made to ensure accuracy, but the Publisher will not be held responsible for any remaining inaccuracies. If you require any further clarification, please contact our Customer Services Department.