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Article: A new method of culturing and transferring iris pigment epithelium

TitleA new method of culturing and transferring iris pigment epithelium
Authors
Issue Date1997
PublisherAssociation for Research in Vision and Ophthalmology. The Journal's web site is located at http://www.iovs.org
Citation
Investigative Ophthalmology And Visual Science, 1997, v. 38 n. 11, p. 2255-2260 How to Cite?
AbstractPurpose. To optimize a culture technique and transfer iris pigment epithelial (IPE) cells for cellular studies in vitro. Methods. Porcine iris tissues were obtained, and IPE cells were isolated and cultured at high densities by plating them in the form of drops. Spherically shaped structures containing a high concentration of cells were formed after 7 to 10 days of culture. Cells were subcultured by transferring spheres to new culture dishes without employing enzymatic dissociation. The purity of IPE cells was determined by pigmentation and cytokeratin labeling. Proliferation was assessed by incorporation of 5-bromo-2'-deoxyuridine. Cellular structure was analyzed under the light and electron microscopes and function was assayed by rod outer segment phagocytosis. Results. Iris pigment epithelial cells, when cultured at high densities, tended to form elevated spherical structures containing viable cells. The cultured cells were pigmented and showed positive labeling with a monoclonal cytokeratin antibody. The IPE cells proliferated and migrated from the spheres to form monolayers. Cells originating from the transferred spheres also continued to proliferate and to migrate in a similar manner to the originally cultivated cells to form monolayers after 7 to 10 days. These cells were able to phagocytose rod outer segments. Conclusions. This new method provides a simple method of culturing a large quantity of IPE cells. The high yield of pure IPE cells and the ease of transfer provide an ideal means to study them at the cellular level.
Persistent Identifierhttp://hdl.handle.net/10722/176346
ISSN
2015 Impact Factor: 3.427
2015 SCImago Journal Rankings: 2.008
ISI Accession Number ID
References

 

DC FieldValueLanguage
dc.contributor.authorRezai, KAen_US
dc.contributor.authorLai, WWen_US
dc.contributor.authorFarrokhSiar, Len_US
dc.contributor.authorPearlman, Jen_US
dc.contributor.authorShu, Jen_US
dc.contributor.authorPatel, SCen_US
dc.contributor.authorErnest, JTen_US
dc.date.accessioned2012-11-26T09:10:43Z-
dc.date.available2012-11-26T09:10:43Z-
dc.date.issued1997en_US
dc.identifier.citationInvestigative Ophthalmology And Visual Science, 1997, v. 38 n. 11, p. 2255-2260en_US
dc.identifier.issn0146-0404en_US
dc.identifier.urihttp://hdl.handle.net/10722/176346-
dc.description.abstractPurpose. To optimize a culture technique and transfer iris pigment epithelial (IPE) cells for cellular studies in vitro. Methods. Porcine iris tissues were obtained, and IPE cells were isolated and cultured at high densities by plating them in the form of drops. Spherically shaped structures containing a high concentration of cells were formed after 7 to 10 days of culture. Cells were subcultured by transferring spheres to new culture dishes without employing enzymatic dissociation. The purity of IPE cells was determined by pigmentation and cytokeratin labeling. Proliferation was assessed by incorporation of 5-bromo-2'-deoxyuridine. Cellular structure was analyzed under the light and electron microscopes and function was assayed by rod outer segment phagocytosis. Results. Iris pigment epithelial cells, when cultured at high densities, tended to form elevated spherical structures containing viable cells. The cultured cells were pigmented and showed positive labeling with a monoclonal cytokeratin antibody. The IPE cells proliferated and migrated from the spheres to form monolayers. Cells originating from the transferred spheres also continued to proliferate and to migrate in a similar manner to the originally cultivated cells to form monolayers after 7 to 10 days. These cells were able to phagocytose rod outer segments. Conclusions. This new method provides a simple method of culturing a large quantity of IPE cells. The high yield of pure IPE cells and the ease of transfer provide an ideal means to study them at the cellular level.en_US
dc.languageengen_US
dc.publisherAssociation for Research in Vision and Ophthalmology. The Journal's web site is located at http://www.iovs.orgen_US
dc.relation.ispartofInvestigative Ophthalmology and Visual Scienceen_US
dc.subject.meshAnimalsen_US
dc.subject.meshCell Culture Techniques - Methodsen_US
dc.subject.meshCell Divisionen_US
dc.subject.meshCell Movementen_US
dc.subject.meshCell Survivalen_US
dc.subject.meshDna - Biosynthesisen_US
dc.subject.meshDna Replicationen_US
dc.subject.meshIris - Cytology - Physiologyen_US
dc.subject.meshPhagocytosis - Physiologyen_US
dc.subject.meshPigment Epithelium Of Eye - Cytology - Physiologyen_US
dc.subject.meshSwineen_US
dc.titleA new method of culturing and transferring iris pigment epitheliumen_US
dc.typeArticleen_US
dc.identifier.emailLai, WW: wicolai@hku.hken_US
dc.identifier.authorityLai, WW=rp00531en_US
dc.description.naturelink_to_OA_fulltexten_US
dc.identifier.pmid9344348-
dc.identifier.scopuseid_2-s2.0-0030749107en_US
dc.relation.referenceshttp://www.scopus.com/mlt/select.url?eid=2-s2.0-0030749107&selection=ref&src=s&origin=recordpageen_US
dc.identifier.volume38en_US
dc.identifier.issue11en_US
dc.identifier.spage2255en_US
dc.identifier.epage2260en_US
dc.identifier.isiWOS:A1997YC22800010-
dc.publisher.placeUnited Statesen_US
dc.identifier.scopusauthoridRezai, KA=7004959843en_US
dc.identifier.scopusauthoridLai, WW=7402231098en_US
dc.identifier.scopusauthoridFarrokhSiar, L=6602082268en_US
dc.identifier.scopusauthoridPearlman, J=7006908369en_US
dc.identifier.scopusauthoridShu, J=7102476894en_US
dc.identifier.scopusauthoridPatel, SC=7403903292en_US
dc.identifier.scopusauthoridErnest, JT=7006438134en_US

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