The use of fluorescence microscopy to define polymer localisation to the late endocytic compartments in cells that are targets for drug delivery

Simon C.W. Richardson; Kerri-Lee Wallom; Elaine L. Ferguson; Samuel P.E. Deacon; Matthew W. Davies; Alison J. Powell; Robert C. Piper; Ruth Duncan

doi:10.1016/j.jconrel.2007.12.015

Back

The use of fluorescence microscopy to define polymer localisation to the late endocytic compartments in cells that are targets for drug delivery

Journal article

Peer reviewed

The use of fluorescence microscopy to define polymer localisation to the late endocytic compartments in cells that are targets for drug delivery

Simon C.W. Richardson, Kerri-Lee Wallom, Elaine L. Ferguson, Samuel P.E. Deacon, Matthew W. Davies, Alison J. Powell, Robert C. Piper and Ruth Duncan

Journal of controlled release, Vol.127(1), pp.1-11

2008

DOI: 10.1016/j.jconrel.2007.12.015

PMCID: PMC6661902

PMID: 18281120

Files and links (1)

url

https://www.ncbi.nlm.nih.gov/pmc/articles/6661902View

Open Access

Abstract

Macromolecular therapeutics and nano-sized drug delivery systems often require localisation to specific intracellular compartments. In particular, efficient endosomal escape, retrograde trafficking, or late endocytic/lysosomal activation are often prerequisites for pharmacological activity. The aim of this study was to define a fluorescence microscopy technique able to confirm the localisation of water-soluble polymeric carriers to late endocytic intracellular compartments. Three polymeric carriers of different molecular weight and character were studied: dextrin (Mw ~ 50,000 g/mol), a N-(2-hydroxypropyl)methacrylamide (HPMA) copolymer (Mw ~ 35,000 g/mol) and polyethylene glycol (PEG) (Mw 5000 g/mol). They were labelled with Oregon Green (OG) (0.3–3 wt.%; < 3% free OG in respect of total). A panel of relevant target cells were used: THP-1, ARPE-19, and MCF-7 cells, and primary bovine chondrocytes (currently being used to evaluate novel polymer therapeutics) as well as NRK and Vero cells as reference controls. Specific intracellular compartments were marked using either endocytosed physiological standards, Marine Blue (MB) or Texas-red (TxR)-Wheat germ agglutinin (WGA), TxR-Bovine Serum Albumin (BSA), TxR-dextran, ricin holotoxin, C6-7-nitro-2,1,3-benzoxadiazol-4-yl (NBD)-labelled ceramide and TxR-shiga toxin B chain, or post-fixation immuno-staining for early endosomal antigen 1 (EEA1), lysosomal-associated membrane proteins (LAMP-1, Lgp-120 or CD63) or the Golgi marker GM130. Co-localisation with polymer–OG conjugates confirmed transfer to discreet, late endocytic (including lysosomal) compartments in all cells types. The technique described here is a particularly powerful tool as it circumvents fixation artefacts ensuring the retention of water-soluble polymers within the vesicles they occupy.

Endocytosis

Intracellular trafficking

Nanomedicine

Polymer conjugates

Details

Title: Subtitle: The use of fluorescence microscopy to define polymer localisation to the late endocytic compartments in cells that are targets for drug delivery
Creators: Simon C.W. Richardson - Cardiff University
Kerri-Lee Wallom - Cardiff University
Elaine L. Ferguson - Cardiff University
Samuel P.E. Deacon - Centre for Polymer Therapeutics, Welsh School of Pharmacy, Cardiff University, Redwood Building, King Edward VII Av, Cardiff, Wales CF10 3XF, UK
Matthew W. Davies - Cardiff University
Alison J. Powell - Cardiff University
Robert C. Piper - University of Iowa
Ruth Duncan - Cardiff University
Resource Type: Journal article
Publication Details: Journal of controlled release, Vol.127(1), pp.1-11
DOI: 10.1016/j.jconrel.2007.12.015
PMID: 18281120
PMCID: PMC6661902
NLM abbreviation: J Control Release
ISSN: 0168-3659
eISSN: 1873-4995
Publisher: Elsevier B.V
Language: English
Date published: 2008
Academic Unit: Molecular Physiology and Biophysics; Medicine Administration; Internal Medicine
Record Identifier: 9984297609602771

Metrics

10 Record Views

52 Times Cited - Web of Science