Phenotypic shift of human amniotic epithelial cells in culture is associated with reduced osteogenic differentiation in vitro

Background Amniotic membrane is a highly promising cell source for tissue engineering. Being part of the placenta, this tissue is abundantly available. It can be processed easily to yield large amounts of epithelial and mesenchymal cells that have shown broad differentiation potential. For tissue-en...

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Veröffentlicht in:Cytotherapy (Oxford, England) England), 2008, Vol.10 (7), p.743-752
Hauptverfasser: Stadler, G, Hennerbichler, S, Lindenmair, A, Peterbauer, A, Hofer, K, van Griensven, M, Gabriel, C, Redl, H, Wolbank, S
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container_end_page 752
container_issue 7
container_start_page 743
container_title Cytotherapy (Oxford, England)
container_volume 10
creator Stadler, G
Hennerbichler, S
Lindenmair, A
Peterbauer, A
Hofer, K
van Griensven, M
Gabriel, C
Redl, H
Wolbank, S
description Background Amniotic membrane is a highly promising cell source for tissue engineering. Being part of the placenta, this tissue is abundantly available. It can be processed easily to yield large amounts of epithelial and mesenchymal cells that have shown broad differentiation potential. For tissue-engineering purposes, cells may be applied either directly after isolation from the tissue or after a period of in vitro expansion to obtain higher cell numbers. In order to investigate the advantages and drawbacks of these strategies we compared freshly isolated and cultivated human amniotic epithelial cells (hAEC) regarding their surface antigen (Ag) expression profile and osteogenic differentiation capacity. Methods Expression of surface Ag that are characteristic for mesenchymal stromal and embryonic stem cells was analyzed by flow cytometry. Different protocols for osteogenic and adipogenic differentiation were compared. Results We have demonstrated that expression of surface Ag changes dramatically during cultivation of hAEC. While not or only weakly expressed on primary isolates, the mesenchymal markers CD13, CD44, CD49e, CD54, CD90 and CD105 are strongly up-regulated during in vitro propagation. In contrast, expression of the embryonic markers TRA-1-60 and TRA-1-81, but not SSEA-4, rapidly decreases upon cultivation. This phenotypic shift is associated with a reduction in osteogenic differentiation. Discussion Our results suggest that phenotypic alterations of hAEC during in vitro cultivation might be responsible for a functional reduction of the differentiation potential, which has to be considered for the potential application of these cells in regenerative medicine.
doi_str_mv 10.1080/14653240802345804
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Being part of the placenta, this tissue is abundantly available. It can be processed easily to yield large amounts of epithelial and mesenchymal cells that have shown broad differentiation potential. For tissue-engineering purposes, cells may be applied either directly after isolation from the tissue or after a period of in vitro expansion to obtain higher cell numbers. In order to investigate the advantages and drawbacks of these strategies we compared freshly isolated and cultivated human amniotic epithelial cells (hAEC) regarding their surface antigen (Ag) expression profile and osteogenic differentiation capacity. Methods Expression of surface Ag that are characteristic for mesenchymal stromal and embryonic stem cells was analyzed by flow cytometry. Different protocols for osteogenic and adipogenic differentiation were compared. Results We have demonstrated that expression of surface Ag changes dramatically during cultivation of hAEC. While not or only weakly expressed on primary isolates, the mesenchymal markers CD13, CD44, CD49e, CD54, CD90 and CD105 are strongly up-regulated during in vitro propagation. In contrast, expression of the embryonic markers TRA-1-60 and TRA-1-81, but not SSEA-4, rapidly decreases upon cultivation. This phenotypic shift is associated with a reduction in osteogenic differentiation. Discussion Our results suggest that phenotypic alterations of hAEC during in vitro cultivation might be responsible for a functional reduction of the differentiation potential, which has to be considered for the potential application of these cells in regenerative medicine.</description><identifier>ISSN: 1465-3249</identifier><identifier>EISSN: 1477-2566</identifier><identifier>DOI: 10.1080/14653240802345804</identifier><identifier>PMID: 18985480</identifier><language>eng</language><publisher>England: Elsevier Inc</publisher><subject>Advanced Basic Science ; Amnion - cytology ; Amnion - physiology ; Cell Culture Techniques ; Cell Differentiation ; Epithelial Cells - cytology ; Epithelial Cells - physiology ; Female ; human amniotic cells ; Humans ; immunophenotype ; Immunophenotyping ; mesenchymal markers ; Mesenchymal Stromal Cells - cytology ; Mesenchymal Stromal Cells - physiology ; Osteogenesis ; Other ; Pregnancy ; stem cell markers</subject><ispartof>Cytotherapy (Oxford, England), 2008, Vol.10 (7), p.743-752</ispartof><rights>International Society for Cellular Therapy</rights><rights>2008 International Society for Cellular Therapy</rights><rights>2008 Informa UK Ltd All rights reserved: reproduction in whole or part not permitted 2008</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c489t-7eb8b729799d85a45095aa38e8ae963d9224dace1460959d1c8f7b62ae6289913</citedby><cites>FETCH-LOGICAL-c489t-7eb8b729799d85a45095aa38e8ae963d9224dace1460959d1c8f7b62ae6289913</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.tandfonline.com/doi/pdf/10.1080/14653240802345804$$EPDF$$P50$$Ginformahealthcare$$H</linktopdf><linktohtml>$$Uhttps://www.tandfonline.com/doi/full/10.1080/14653240802345804$$EHTML$$P50$$Ginformahealthcare$$H</linktohtml><link.rule.ids>314,780,784,4024,27923,27924,27925,61221,61402</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/18985480$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Stadler, G</creatorcontrib><creatorcontrib>Hennerbichler, S</creatorcontrib><creatorcontrib>Lindenmair, A</creatorcontrib><creatorcontrib>Peterbauer, A</creatorcontrib><creatorcontrib>Hofer, K</creatorcontrib><creatorcontrib>van Griensven, M</creatorcontrib><creatorcontrib>Gabriel, C</creatorcontrib><creatorcontrib>Redl, H</creatorcontrib><creatorcontrib>Wolbank, S</creatorcontrib><title>Phenotypic shift of human amniotic epithelial cells in culture is associated with reduced osteogenic differentiation in vitro</title><title>Cytotherapy (Oxford, England)</title><addtitle>Cytotherapy</addtitle><description>Background Amniotic membrane is a highly promising cell source for tissue engineering. Being part of the placenta, this tissue is abundantly available. It can be processed easily to yield large amounts of epithelial and mesenchymal cells that have shown broad differentiation potential. For tissue-engineering purposes, cells may be applied either directly after isolation from the tissue or after a period of in vitro expansion to obtain higher cell numbers. In order to investigate the advantages and drawbacks of these strategies we compared freshly isolated and cultivated human amniotic epithelial cells (hAEC) regarding their surface antigen (Ag) expression profile and osteogenic differentiation capacity. Methods Expression of surface Ag that are characteristic for mesenchymal stromal and embryonic stem cells was analyzed by flow cytometry. Different protocols for osteogenic and adipogenic differentiation were compared. Results We have demonstrated that expression of surface Ag changes dramatically during cultivation of hAEC. While not or only weakly expressed on primary isolates, the mesenchymal markers CD13, CD44, CD49e, CD54, CD90 and CD105 are strongly up-regulated during in vitro propagation. In contrast, expression of the embryonic markers TRA-1-60 and TRA-1-81, but not SSEA-4, rapidly decreases upon cultivation. This phenotypic shift is associated with a reduction in osteogenic differentiation. 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subjects Advanced Basic Science
Amnion - cytology
Amnion - physiology
Cell Culture Techniques
Cell Differentiation
Epithelial Cells - cytology
Epithelial Cells - physiology
Female
human amniotic cells
Humans
immunophenotype
Immunophenotyping
mesenchymal markers
Mesenchymal Stromal Cells - cytology
Mesenchymal Stromal Cells - physiology
Osteogenesis
Other
Pregnancy
stem cell markers
title Phenotypic shift of human amniotic epithelial cells in culture is associated with reduced osteogenic differentiation in vitro
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