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Attachment and viability of arpe-19 cells on biodegradable polymer microspheres

Attachment and viability of arpe-19 cells on biodegradable polymer microspheres
Attachment and viability of arpe-19 cells on biodegradable polymer microspheres
Purpose: the retinal pigment epithelium is a primary target for cellular replacement in degenerative ocular diseases such as age related macular degeneration. Transplantation of cells to the subretinal space is typically achieved by injection of cell suspensions, however there are a number of disadvantages associated with this method, including cell death and incorrectly localised or disorganised grafts. Recent studies have demonstrated that growth of retinal cells on polymer scaffolds can facilitate the transplantation of cells, which may reduce cell death by up to 10 fold (1). There have only been limited investigations to determine the most appropriate polymer composition and conformation for optimal cell delivery.

Methods: using a technique adapted from (2). We manufactured biodegradable microspheres using five different blends of poly(L-lactic acid) (PLLA) with poly(D, L-lactic-glycolic acid) (PLGA). Scanning electron microscopy (SEM) was used to characterise the microsphere size range produced and surface characteristics. Microsphere surface chemistry was modified using the extracellular matrix protein laminin. The microsphere blends were seeded with an ARPE-19 cell line and maintained in culture for up to 2 weeks. Cell adherence, proliferation and phenotype were assessed by immunocytochemistry using antisera directed against proliferating cell nuclear antigen, RPE65 and pan-cytokeratin. Cell survival was quantified by measurement of apoptosis and cell membrane integrity.

Results: microsphere diameter and surface characteristics were dependent on the copolymer blend ratios and concentration of copolymer used. Attachment of ARPE-19 cells to coated and uncoated microsperes was confirmed. Cells proliferated, remained viable and retained phenotypic characteristics on all blends. There was a trends toward an increase in cell death on microspheres predominantely comprised of PLGA, however this may be attributed to the smaller size of these spheres leading to cell clumping, rather than a cytotoxic effect.

Conclusions: from this study we found microspheres to be an efficacious scaffold configuration for cell attachment and growth in vitro with preservation of phenotypic characteristics associated with minimal demonstrable cytotoxicity.(1) Tomita M et al. Stem Cells 2005 10:1579-88.(2) Gabler F et al. Biomolecular Engineering 2007 24: 515-520
5243
Thomson, Heather Anne Jane
7db0d6dd-4e3c-47b0-a281-9a04c0b70252
Treharne, Andrew John
a158bb09-9d41-4b3a-ae02-ccbd76dd9417
Grossel, Martin Christopher
403bf3ff-6364-44e9-ab46-52d84c6f0d56
Lotery, Andrew John
5ecc2d2d-d0b4-468f-ad2c-df7156f8e514
Thomson, Heather Anne Jane
7db0d6dd-4e3c-47b0-a281-9a04c0b70252
Treharne, Andrew John
a158bb09-9d41-4b3a-ae02-ccbd76dd9417
Grossel, Martin Christopher
403bf3ff-6364-44e9-ab46-52d84c6f0d56
Lotery, Andrew John
5ecc2d2d-d0b4-468f-ad2c-df7156f8e514

Thomson, Heather Anne Jane, Treharne, Andrew John, Grossel, Martin Christopher and Lotery, Andrew John (2010) Attachment and viability of arpe-19 cells on biodegradable polymer microspheres. ARVO Meeting Abstracts, 51 (5), 5243.

Record type: Article

Abstract

Purpose: the retinal pigment epithelium is a primary target for cellular replacement in degenerative ocular diseases such as age related macular degeneration. Transplantation of cells to the subretinal space is typically achieved by injection of cell suspensions, however there are a number of disadvantages associated with this method, including cell death and incorrectly localised or disorganised grafts. Recent studies have demonstrated that growth of retinal cells on polymer scaffolds can facilitate the transplantation of cells, which may reduce cell death by up to 10 fold (1). There have only been limited investigations to determine the most appropriate polymer composition and conformation for optimal cell delivery.

Methods: using a technique adapted from (2). We manufactured biodegradable microspheres using five different blends of poly(L-lactic acid) (PLLA) with poly(D, L-lactic-glycolic acid) (PLGA). Scanning electron microscopy (SEM) was used to characterise the microsphere size range produced and surface characteristics. Microsphere surface chemistry was modified using the extracellular matrix protein laminin. The microsphere blends were seeded with an ARPE-19 cell line and maintained in culture for up to 2 weeks. Cell adherence, proliferation and phenotype were assessed by immunocytochemistry using antisera directed against proliferating cell nuclear antigen, RPE65 and pan-cytokeratin. Cell survival was quantified by measurement of apoptosis and cell membrane integrity.

Results: microsphere diameter and surface characteristics were dependent on the copolymer blend ratios and concentration of copolymer used. Attachment of ARPE-19 cells to coated and uncoated microsperes was confirmed. Cells proliferated, remained viable and retained phenotypic characteristics on all blends. There was a trends toward an increase in cell death on microspheres predominantely comprised of PLGA, however this may be attributed to the smaller size of these spheres leading to cell clumping, rather than a cytotoxic effect.

Conclusions: from this study we found microspheres to be an efficacious scaffold configuration for cell attachment and growth in vitro with preservation of phenotypic characteristics associated with minimal demonstrable cytotoxicity.(1) Tomita M et al. Stem Cells 2005 10:1579-88.(2) Gabler F et al. Biomolecular Engineering 2007 24: 515-520

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More information

Published date: May 2010
Venue - Dates: The Association for Research in Vision and Ophthalmology Conference, Fort Lauderdale, United States, 2010-04-30

Identifiers

Local EPrints ID: 181043
URI: http://eprints.soton.ac.uk/id/eprint/181043
PURE UUID: 2b2e0ca3-c516-434f-b2f8-7047791504e1
ORCID for Martin Christopher Grossel: ORCID iD orcid.org/0000-0001-7469-6854
ORCID for Andrew John Lotery: ORCID iD orcid.org/0000-0001-5541-4305

Catalogue record

Date deposited: 15 Apr 2011 08:43
Last modified: 30 Jan 2020 01:32

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