REVIEW PAPER
Cell-based assays in high-throughput mode (HTS)
1
Universite du Quebec a Montreal, Department of Biological Sciences, Montreal, Canada
Submission date: 2016-04-18
Final revision date: 2016-06-28
Acceptance date: 2016-08-02
Publication date: 2016-11-02
BioTechnologia 2016;97(3):227-234
KEYWORDS
TOPICS
ABSTRACT
Typically, novel compounds are screened by the millions, a process known as high-throughput screening (HTS). HTS allows for the screening of millions of potential drugs in a relatively short period of time. All compounds are initially subjected to various tests to determine safety and efficacy. At the molecular level, typically two types of tests are available: in vitro and cell-based assays (i.e., in vivo ). The distinction between a cell-based assay and an in vitro screening is that the cell-based assay utilizes live cells – approximately 50 000 cells are seeded onto the floor of the well. Cell-based assays are used to measure proliferation, toxicity, marker production, motility, activation of signaling pathways, and changes in morphology. In such cases, other factors such as 2D versus 3D culture or static versus profusion cultures might also contribute to the results obtained. This study emphasizes the positive aspects of using cell-based assays in high-throughput mode.
REFERENCES (43)
1.
Astashkina A., Mann B., Grainger D. (2012) A critical evaluation of in vitro cell culture models for high-throughput drug screening and toxicity. Pharmacol. Ther. 134: 82-106.
2.
Bedard J., May S., Barbeau D. et al. (1999) A high throughput colorimetric cell proliferation assay for the identification of human cytomegalovirus inhibitors. Antiviral Res. 41: 35-43.
3.
Bray M.-A., Carpenter A. (2013) Advanced assay development guidelines for image-based high content screening and analysis. [in:] Assay Guidance Manual [Internet]. Ed. Sittampalam G.S., Coussens N.P., Nelson H. et al., Bethesda (MD): Eli Lilly & Company and the National Center for Advancing Translational Sciences. http: www.ncbi. nlm.nih.gov/books/NBK126174/.
4.
Breier J., Radio N., Mundy W., Shafer T. (2008) Development of a high-throughput screening assay for chemical effects on proliferation and viability of immortalized human neural progenitor cells. Toxicol. Sci. 105: 119-133.
5.
Chattopadhyay A. (2014) GPCRs: lipid-dependent membrane receptors that act as drug targets. Adv. Biol. 2014: ID 143023.
6.
Corning Corning Microplates Product Selection Guide For Assays and Drug Discovery.
7.
Cottet M., Faklaris O., Zwier J. et al. (2011) Original fluorescent ligand-based assays open new perspectives in G-protein coupled receptor drug screening. Pharmaceuticals 4: 202-214.
8.
Du G., Fang Q., den Toonder J.M.J. (2016) Microfluidics for cell-based high throughput screening platforms: A review. Anal. Chim. Acta 903: 36-50.
9.
Edmondson R., Broglie J.J., Adcock A.F., Yang L. (2014) Three-dimensional cell culture systems and their applications in drug discovery and cell-based biosensors. Assay Drug Dev. Technol. 12: 207-218.
10.
Fang Y., Frutos A., Verklereen R. (2008) Label-free cell-based assays for GPCR screening. Comb. Chem. High Throughput Screen 11: 357-369.
11.
Fennema E., Rivron N., Rouwkema J. et al. (2013) Spheroid culture as a tool for creating 3D complex tissues. Trends Biotechnol. 31: 108-115. doi: 10.1016/j.tibtech.2012.12.003.
12.
Fox S., Farr-Jones S., Sopchak L. et al. (2006) High-throughput screening: update on practices and success. J. Biomol. Screen 11: 864-869.
13.
Frank An W., Tolliday N. (2010) Cell-based assays for highthroughput screening. Mol. Biotechnol. 45: 180-186.
14.
Garbison K., Heinz B., Lajiness M. et al. (2012) Impedance-based technologies. [in:] Assay Guidance Manual [Internet]. Ed. Sittampalam G.S., Coussens N.P., Nelson H. et al., Bethesda (MD): Eli Lilly & Company and the National Center for Advancing Translational Sciences. http: www. ncbi.nlm.nih.gov/ books/NBK91999/.
15.
Gibbs J. (2001) Selecting the detection system – colorimetric, fluorescent, luminescent methods. ELISA Technical Bulletin.
16.
Grepin C., Pernelle C. (2000) High-throughput screening evolution of homogeneous time resolved fluorescence (HTRF) technology for HTS. DDT 5: 212-214.
17.
Gribbon P., Sewing A. (2003) Fluorescence readouts in HTS: no gain without pain? Drug Discov. Today 8: 1035-1043.
18.
Johnston P., Johnston P. (2002) Cellular platforms for HTS: three case studies. Drug Discov. Today 7: 353-363.
19.
Justice B.A., Badr N.A., Felder R.A. (2009) 3D cell culture opens new dimensions in cell-based assays. Drug Discov. Today 14: 102-107.
20.
Kain S. (1999) Green fluorescent protein (GFP): applications in cell-based assays for drug discovery. Drug Discov. Today 4: 304-312.
21.
Kim L., Toh Y., Voldman J., Yu H. (2007) A practical guide to microfluidic perfusion culture of adherent mammalian cells. Lab Chip 7: 681-694.
22.
Lodish H., Berk A., Zipursky S. (2000) Section 20.3G potein – coupled receptors and their effectors. [in:] Molecular Cell Biology. Ed. W.H. Freeman and Company, New York.
23.
Macarron R., Banks M.N., Bojanic D. et al. (2011) Impact of high-throughput screening in biomedical research. Nat. Rev. Drug Discov. 10: 188-195.
24.
Mcguinness R. (2007) Impedance-based cellular assay technologies: recent advances, future promise. Curr. Opin. Pharmacol. 7: 535-540.
25.
McKim J.J. (2010) Building a tiered approach to in vitro predictive toxicity screening: a focus on assays with in vivo relevance. Comb. Chem. High Throughput Screen. 13: 188-206.
26.
Michelini E., Cevenini L., Mezzanotte L. et al. (2010) Cell-based assays: fuelling drug discovery. Anal. Bioanal. Chem. 398: 227-222.
27.
Papadimitriou E., Lelkes P. (1993) Measurement of cell numbers in microtiter culture plates using the fluorescent dye Hoechst 33258. J. Immunol. Meth. 162: 41-45.
28.
Peterson K.R., Costa F.C., Fedosyuk H. et al. (2014) A cellbased high-throughput screen for novel chemical inducers of fetal hemoglobin for treatment of hemoglobinopathies. PLoS ONE 9: e107006. doi: 10.1371/journal.pone. 0107006.
29.
Riss T., Moravec R., Niles A. et al. (2013) Cell viability assays. [in:] Assay Guidance Manual [Internet]. Ed. Sittampalam G.S., Coussens N.P., Nelson H. et al., Bethesda (MD): Eli Lilly & Company and the National Center for Advancing Translational Sciences.
30.
Rogers M. (1997) Light on high-throughput screening: fluorescence- based assay technologies. Drug Discov. Today 2: 156-160.
31.
Schoonen W., Westerink W., Horbach G. (2009) High-throughput screening for analysis of in vitro toxicity. [in:] Molecular, Clinical and Environmental Toxicology. Birkhäuser Basel: 401-452.
32.
Slater K. (2001) Cytotoxicity tests for high-throughput drug discovery. Curr. Opin. Biotechnol. 12: 70-74.
33.
Smitskamp-Wilms E., Pinedo H., Veerman G. et al. (1998) Postconfluent multilayered cell line cultures for selective screening of gemcitabine. Eur. J. Cancer 34: 921-926.
34.
Szymański P., Markowicz M., Mikiciuk-Olasik E. (2011) Adaptation of high-throughput screening in drug discovery – toxicological screening tests. Int. J. Mol. Sci. 13: 427- 452. doi: 10.3390/ijms13010427.
35.
Takakura H., Hattori M., Tanaka M., Ozawa T. (2015) Cell-based assays and animal models for GPCR drug screening. [in:] G protein-coupled receptor screening assays: methods and protocols. Humana Press, New York, Springer: 257-270.
36.
Tautermann C.S. (2014) GPCR structures in drug design, emerging opportunities with new structures. Bioorg. Med. Chem. Lett. 24: 4073-4079.
37.
Thomsen W., Frazer J., Unett D. (2005) Functional assays for screening GPCR targets. Curr. Opin. Biotechnol. 16: 655- 665.
38.
Tung Y., Hsiao A., Allen S. et al. (2011) High-throughput 3D spheroid culture and drug testing using a 384 hanging drop array. Analyst 136: 473-478.
39.
Yeon J., Park J. (2005) Cytotoxicity test based on electrochemical impedance measurement of HepG2 cultured in microfabricated cell chip. Anal. Biochem. 341: 308-315.
40.
Yu H., Chung H., Yu Y. et al. (2003) A new rapid and nonradioactive assay for monitoring and determining the proliferation of retinal pigment epithelial cells. Korean J. Ophthalmol. 17: 29.
41.
Zang R., Li D., Tang I. et al. (2012) Cell-based assays in highthroughput screening for drug discovery. Int. J. Biotechnol. Wellness Ind. 1: 31-51.
42.
Zhang J., Chung T., Oldenburg K. (1999) A simple statistical parameter for use in evaluation and validation of high throughput screening assays. J. Biomol. Screen 4: 67-73.
43.
Zhang Z., Guan N., Li T. et al. (2012) Quality control of cellbased high-throughput drug screening. Acta Pharm. Sin. B2: 429-438.
| eISSN: | 2353-9461 |
| ISSN: | 0860-7796 |
We process personal data collected when visiting the website. The function of obtaining information about users and their behavior is carried out by voluntarily entered information in forms and saving cookies in end devices. Data, including cookies, are used to provide services, improve the user experience and to analyze the traffic in accordance with the Privacy policy. Data are also collected and processed by Google Analytics tool (more).
You can change cookies settings in your browser. Restricted use of cookies in the browser configuration may affect some functionalities of the website.
You can change cookies settings in your browser. Restricted use of cookies in the browser configuration may affect some functionalities of the website.
