Abstract
The most critical stage in evaluation of a novel drug or its delivery system is assessment of its safety and efficacy. Traditionally, in vivo animal models were used for this assessment. However, due to growing ethical concerns in animal usage, these in vivo animal models have largely been replaced by cell-based assays. Cell-based assays offer several advantages which have been described in this section. This chapter describes in detail the protocols along with critical parameters for various cell-based assays which can be used for evaluation of targeted drug delivery systems for cancer and infectious diseases.
Aakruti Kaikini and Vaibhavi Peshattiwar have equally contributed to this work.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Abbreviations
- 3H-T:
-
3H-labeled thymidine
- ATCC:
-
American Type Culture Collection
- bFGF:
-
Basic fibroblast growth factor
- BrdU:
-
5-Bromo-2′-deoxyuridine
- CAM:
-
Chick embryo chorioallantoic membrane
- cfu:
-
Colony-forming units
- CLSI:
-
The Clinical and Laboratory Standards Institute
- DMSO:
-
Dimethyl sulfoxide
- DTT:
-
Dithiothreitol
- EBM:
-
Endothelial basal media
- EDTA:
-
Ethylenediaminetetraacetic acid
- EGTA:
-
(Ethylene glycol-bis(β-aminoethyl ether)-N,N,N′,N′-tetraacetic acid)
- ESCMID:
-
European Society of Clinical Microbiology and Infectious Diseases
- FITC:
-
Fluorescein isothiocyanate
- FRET:
-
Forster resonance energy transfer
- HPLC:
-
High performance liquid chromatography
- HUVECs:
-
Human umbilical vein endothelial cells
- LC-MS:
-
Liquid chromatography-mass spectrometry
- MHB:
-
Mueller–Hinton broth
- MIC:
-
Minimum inhibitory concentration
- MTT:
-
(3-[4,5-Dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide)
- NDDS:
-
Novel drug delivery system
- PAE:
-
Post antibiotic effect
- PI:
-
Propidium iodide
- UV:
-
Ultraviolet
- VEGF:
-
Vascular endothelial growth factor
References
Hughes JP, Rees S, Kalindjian SB, Philpott KL. Principles of early drug discovery. Br J Pharmacol. 2011;162(6):1239–49.
Goldberg AM. Animals and alternatives: societal expectations and scientific need. Altern Lab Anim. 2004;32(6):545–51.
Feng Y, Mitchison TJ, Bender A, Young DW, Tallarico JA. Multi-parameter phenotypic profiling: using cellular effects to characterize small-molecule compounds. Nat Rev Drug Discov. 2009;8(7):567–78.
Clemons PA. Complex phenotypic assays in high-throughput screening. Curr Opin Chem Biol. 2004;8(3):334–8.
van Meerloo JKG, Cloos J. Cell sensitivity assays: the MTT assay. Methods Mol Biol. 2011;731:237–45.
Romar GA, Kupper TS, Divito SJ. Research techniques made simple: techniques to assess cell proliferation. J Invest Dermatol. 2016;136(1):e1–7.
Terry NH, White RA. Flow cytometry after bromodeoxyuridine labeling to measure S and G2+M phase durations plus doubling times in vitro and in vivo. Nat Protoc. 2006;1(2):859–69.
Nguyen M, Shing Y, Folkman J. Quantitation of angiogenesis and antiangiogenesis in the chick embryo chorioallantoic membrane. Microvasc Res. 1994;47(1):31–40.
Hlushchuk R, Bronnimann D, Correa Shokiche C, Schaad L, Triet R, Jazwinska A, et al. Zebrafish caudal fin angiogenesis assay-advanced quantitative assessment including 3-way correlative microscopy. PLoS One. 2016;11(3):e0149281.
Cao Z, Zheng L, Zhao J, Zhuang Q, Hong Z, Lin W. Anti-angiogenic effect of Livistona chinensis seed extract in vitro and in vivo. Oncol Lett. 2017;14(6):7565–70.
Nitiss JL, Soans E, Rogojina A, Seth A, Mishina M. Topoisomerase assays. Curr Protoc Pharmacol. 2012; Chapter 3:Unit 3; https://doi.org/10.1002/0471141755.ph0303s57.
Pozarowski P, Darzynkiewicz Z. Analysis of cell cycle by flow cytometry. Methods Mol Biol. 2004;281:301–11.
Rieger AM, Nelson KL, Konowalchuk JD, Barreda DR. Modified annexin V/propidium iodide apoptosis assay for accurate assessment of cell death. J Vis Exp. 2011;(50) https://doi.org/10.3791/2597.
Rehm M, Parsons MJ, Bouchier-Hayes L. Measuring caspase activity by Forster resonance energy transfer. Cold Spring Harb Protoc. 2015;2015(1):pdb prot082560.
Muller C, Schubiger PA, Schibli R. In vitro and in vivo targeting of different folate receptor-positive cancer cell lines with a novel 99mTc-radiofolate tracer. Eur J Nucl Med Mol Imaging. 2006;33(10):1162–70.
EUCAST. European Committee for Antimicrobial Susceptibility Testing (EUCAST). Clin Microbiol Infect. 2003;9(8):9–15.
Wayne P, CLSI. Performance standards for antimicrobial susceptibility testing. CLSI supplement M100. 29th ed. Wayne: Clinical and Laboratory Standards Institute; 2019.
Wiegand I, Hilpert K, Hancock RE. Agar and broth dilution methods to determine the minimal inhibitory concentration (MIC) of antimicrobial substances. Nat Protoc. 2008;3(2):163–75.
Forry SP, Madonna MC, Lopez-Perez D, Lin NJ, Pasco MD. Automation of antimicrobial activity screening. AMB Express. 2016;6(1):20.
Zhao DH, Yu Y, Zhou YF, Shi W, Deng H, Liu YH. Postantibiotic effect and postantibiotic sub-minimum inhibitory concentration effect of valnemulin against Staphylococcus aureus isolates from swine and chickens. Lett Appl Microbiol. 2014;58(2):150–5.
Motyl M, Dorso K, Barrett J, Giacobbe R. Basic microbiological techniques used in antibacterial drug discovery. Curr Protoc Pharmacol. 2006; Chapter 13:Unit13A 3; https://doi.org/10.1002/0471141755.ph13a03s31.
Author information
Authors and Affiliations
Corresponding authors
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2019 American Association of Pharmaceutical Scientists
About this chapter
Cite this chapter
Kaikini, A., Peshattiwar, V., Devarajan, P.V., Dandekar, P., Sathaye, S. (2019). Protocols for Cellular Evaluation of Targeted Drug Delivery Systems for Cancer and Infectious Diseases. In: Devarajan, P., Dandekar, P., D'Souza, A. (eds) Targeted Intracellular Drug Delivery by Receptor Mediated Endocytosis. AAPS Advances in the Pharmaceutical Sciences Series, vol 39. Springer, Cham. https://doi.org/10.1007/978-3-030-29168-6_18
Download citation
DOI: https://doi.org/10.1007/978-3-030-29168-6_18
Published:
Publisher Name: Springer, Cham
Print ISBN: 978-3-030-29167-9
Online ISBN: 978-3-030-29168-6
eBook Packages: Biomedical and Life SciencesBiomedical and Life Sciences (R0)