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Article: Isolation of amelogenin-positive ameloblasts from rat mandibular incisor enamel organs by flow cytometry and fluorescence activated cell sorting

TitleIsolation of amelogenin-positive ameloblasts from rat mandibular incisor enamel organs by flow cytometry and fluorescence activated cell sorting
Authors
Issue Date1998
PublisherInforma Healthcare. The Journal's web site is located at http://www.tandf.co.uk/journals/titles/03008207.asp
Citation
Connective Tissue Research, 1998, v. 38 n. 1-4, p. 9-15 How to Cite?
AbstractThe purpose of this study was to use amelogenin as a marker to examine the feasibility of isolating ameloblasts from enamel organ cell populations by fluorescence activated cell sorting. After treating dissected rat enamel organs with proteolytic enzymes to loosen cell attachments and labial connective tissues, dissociated cell suspensions were fixed, then immunostained with rabbit anti-rM179 recombinant amelogenin antibody and FITC-conjugated goat anti-rabbit Ig G antibody. Flow cytometry indicated that about 70% of the total cell sample and virtually all the larger cells therein were amelogenin-positive. Fluorescence activated cell sorting yielded a sample of amelogenin-positive cells at 97% purity. Immunofluorescence microscopy indicated that these isolated amelogenin-positive cells varied widely in size and morphology. This was attributed to loss of intercellular support for ameloblasts once they were dissociated from each other, and to some fragmentation caused when the cells were initially physically removed from the teeth. The results demonstrate that viable ameloblast cell fractions, especially representing cells at the secretory stage, can be purified from enzymic digests of rat enamel organ by sorting on the basis of cell size alone. From these fractions, subpopulations of ameloblasts may be identified when differentiation specific cell surface markers become available.
Persistent Identifierhttp://hdl.handle.net/10722/148134
ISSN
2015 Impact Factor: 1.411
2015 SCImago Journal Rankings: 0.622
ISI Accession Number ID

 

DC FieldValueLanguage
dc.contributor.authorChen, WYen_US
dc.contributor.authorLu, Len_US
dc.contributor.authorMcdonald, Ken_US
dc.contributor.authorOsmond, DGen_US
dc.contributor.authorSmith, CEen_US
dc.date.accessioned2012-05-29T06:11:02Z-
dc.date.available2012-05-29T06:11:02Z-
dc.date.issued1998en_US
dc.identifier.citationConnective Tissue Research, 1998, v. 38 n. 1-4, p. 9-15en_US
dc.identifier.issn0300-8207en_US
dc.identifier.urihttp://hdl.handle.net/10722/148134-
dc.description.abstractThe purpose of this study was to use amelogenin as a marker to examine the feasibility of isolating ameloblasts from enamel organ cell populations by fluorescence activated cell sorting. After treating dissected rat enamel organs with proteolytic enzymes to loosen cell attachments and labial connective tissues, dissociated cell suspensions were fixed, then immunostained with rabbit anti-rM179 recombinant amelogenin antibody and FITC-conjugated goat anti-rabbit Ig G antibody. Flow cytometry indicated that about 70% of the total cell sample and virtually all the larger cells therein were amelogenin-positive. Fluorescence activated cell sorting yielded a sample of amelogenin-positive cells at 97% purity. Immunofluorescence microscopy indicated that these isolated amelogenin-positive cells varied widely in size and morphology. This was attributed to loss of intercellular support for ameloblasts once they were dissociated from each other, and to some fragmentation caused when the cells were initially physically removed from the teeth. The results demonstrate that viable ameloblast cell fractions, especially representing cells at the secretory stage, can be purified from enzymic digests of rat enamel organ by sorting on the basis of cell size alone. From these fractions, subpopulations of ameloblasts may be identified when differentiation specific cell surface markers become available.en_US
dc.languageengen_US
dc.publisherInforma Healthcare. The Journal's web site is located at http://www.tandf.co.uk/journals/titles/03008207.aspen_US
dc.relation.ispartofConnective Tissue Researchen_US
dc.subject.meshAmeloblasts - Cytology - Metabolismen_US
dc.subject.meshAmelogeninen_US
dc.subject.meshAnimalsen_US
dc.subject.meshCell Separation - Methodsen_US
dc.subject.meshDental Enamel Proteins - Immunology - Metabolismen_US
dc.subject.meshDental Pulp - Cytologyen_US
dc.subject.meshEnamel Organ - Cytology - Metabolismen_US
dc.subject.meshFlow Cytometry - Methodsen_US
dc.subject.meshFluorescenceen_US
dc.subject.meshFluorescent Antibody Technique, Indirecten_US
dc.subject.meshIncisor - Cytologyen_US
dc.subject.meshMandibleen_US
dc.subject.meshMicroscopy, Fluorescenceen_US
dc.subject.meshRabbitsen_US
dc.subject.meshRatsen_US
dc.subject.meshRats, Sprague-Dawleyen_US
dc.subject.meshStaining And Labelingen_US
dc.titleIsolation of amelogenin-positive ameloblasts from rat mandibular incisor enamel organs by flow cytometry and fluorescence activated cell sortingen_US
dc.typeArticleen_US
dc.identifier.emailLu, L:liweilu@hkucc.hku.hken_US
dc.identifier.authorityLu, L=rp00477en_US
dc.description.naturelink_to_subscribed_fulltexten_US
dc.identifier.doi10.3109/03008209809017012-
dc.identifier.pmid11063012-
dc.identifier.scopuseid_2-s2.0-0032447940en_US
dc.identifier.volume38en_US
dc.identifier.issue1-4en_US
dc.identifier.spage9en_US
dc.identifier.epage15en_US
dc.identifier.isiWOS:000077394100004-
dc.publisher.placeUnited Kingdomen_US

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