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Conference Paper: Human endometriotic cells exhibit stem-like cell properties

TitleHuman endometriotic cells exhibit stem-like cell properties
Authors
Issue Date2009
PublisherOxford University Press. The Journal's web site is located at http://humrep.oxfordjournals.org/
Citation
The 25th Annual Meeting of the European Society of Human Reproduction and Embryology, Amsterdam, The Netherlands, 28 June – 1 July 2009. In Human Reproduction, 2009, v. 24 n. S1, p. i97 How to Cite?
AbstractIntroduction: Endometriosis, the presence of endometrium outside the uterine cavity, affects 6–10% of women during their reproductive life [1]. It is one of the most common benign gynaecological diseases however, little is known about its pathogenesis. A postulated theory on the aetiology of endometriosis is that viable endometrial cells reach the peritoneal cavity through retrograde menstruation, adhere and form endometriotic lesions. Endometriosis is an estrogen dependent disorder, though the relation between steroids and its pathogenesis remains unclear. Recently stem/progenitor cells have been identified in the endometrium [2, 3]. They exhibit clonogenic activity and have high selfrenewal capability. Therefore, we hypothesize that a reservoir of endometrial epithelial and stromal stem/progenitor cells may contribute to the development of endometriotic lesions. The aims of this study were to 1) determine the clonogenic capacity of human endometrial epithelial and stromal cells from ovarian endometriotic lesions, 2) assess the self renewal capacity of endometriotic clonogenic cells and, 3) examine the cellular composition of the endometriotic clones using phenotypic markers. Materials and Methods: Ovarian endometriotic cysts were obtained from 19 women undergoing laparoscopy. The lesions were dissociated with collagenase to achieve single cell suspensions. Epithelial and stromal cells were separated using Ber-EP4 Dynabeads and cultured at low seeding density (500 cells/cm2) for 21 days. Colonies were stained with hematoxylin to determine the cloning efficiencies (CE) and some individual clones were serially subcloned every 3 –6 weeks to determine self renewal capacity. Immunohistochemistry was performed with: anti-human fibroblast (CD90), anti-human epithelial antigen (Ber-EP4, CD49f), anti-human endometriotic stromal marker (CD10) and estrogen receptor-a (ER-a). Results: Two types of colonies were observed: small loosely packed colonies containing large cells and large colonies with small, densely packed cells. The total CE for epithelial endometriotic cells was 2.92+2.49% (n ¼ 19); 1.70+1.47% for large and 1.22+1.02% for small colonies. Whereas, the total CE for stromal endometriotic cells was 4.67+2.71% (n ¼ 19); 1.54+0.92% for large and 3.13+1.80% for small colonies. There was no significant difference between the two cell types. Single cells derived from large epithelial and stromal endometriotic clones could be subcloned four times more when compared to those from small clones (n ¼ 5). Small epithelial colonies were positive for epithelial marker and negative for the fibroblast marker. However, large colonies were negative for BerEP4 and CD49f suggesting these epithelial markers are lost during in vitro proliferation. Both large and small stromal colonies were positive for CD90 and CD10 and were negative for epithelial markers. Interestingly, both large epithelial and stromal endometriotic clones possessed ER-a immunoreactivity. Conclusions: These results suggest that a small population of cells with stemlike cell properties of clonogenic and self renewal activity may be responsible for the development and progression in endometriosis. References Wheeler, J.M., Epidemiology of endometriosis-associated infertility. J Reprod Med, 1989. 34(1): p. 41–6. Chan, R.W., K.E. Schwab, and C.E. Gargett, Clonogenicity of human endometrial epithelial and stromal cells. Biol Reprod, 2004. 70(6): p. 1738–50. Chan, R.W. and C.E. Gargett, Identification of label-retaining cells in mouse endometrium. Stem Cells, 2006. 24(6): p. 1529–38.
Persistent Identifierhttp://hdl.handle.net/10722/113717
ISSN
2014 Impact Factor: 4.569
2014 SCImago Journal Rankings: 1.972

 

DC FieldValueLanguage
dc.contributor.authorChan, RWS-
dc.contributor.authorNg, EHY-
dc.contributor.authorLee, CKF-
dc.contributor.authorYeung, WSB-
dc.date.accessioned2010-09-26T04:28:07Z-
dc.date.available2010-09-26T04:28:07Z-
dc.date.issued2009-
dc.identifier.citationThe 25th Annual Meeting of the European Society of Human Reproduction and Embryology, Amsterdam, The Netherlands, 28 June – 1 July 2009. In Human Reproduction, 2009, v. 24 n. S1, p. i97-
dc.identifier.issn0268-1161-
dc.identifier.urihttp://hdl.handle.net/10722/113717-
dc.description.abstractIntroduction: Endometriosis, the presence of endometrium outside the uterine cavity, affects 6–10% of women during their reproductive life [1]. It is one of the most common benign gynaecological diseases however, little is known about its pathogenesis. A postulated theory on the aetiology of endometriosis is that viable endometrial cells reach the peritoneal cavity through retrograde menstruation, adhere and form endometriotic lesions. Endometriosis is an estrogen dependent disorder, though the relation between steroids and its pathogenesis remains unclear. Recently stem/progenitor cells have been identified in the endometrium [2, 3]. They exhibit clonogenic activity and have high selfrenewal capability. Therefore, we hypothesize that a reservoir of endometrial epithelial and stromal stem/progenitor cells may contribute to the development of endometriotic lesions. The aims of this study were to 1) determine the clonogenic capacity of human endometrial epithelial and stromal cells from ovarian endometriotic lesions, 2) assess the self renewal capacity of endometriotic clonogenic cells and, 3) examine the cellular composition of the endometriotic clones using phenotypic markers. Materials and Methods: Ovarian endometriotic cysts were obtained from 19 women undergoing laparoscopy. The lesions were dissociated with collagenase to achieve single cell suspensions. Epithelial and stromal cells were separated using Ber-EP4 Dynabeads and cultured at low seeding density (500 cells/cm2) for 21 days. Colonies were stained with hematoxylin to determine the cloning efficiencies (CE) and some individual clones were serially subcloned every 3 –6 weeks to determine self renewal capacity. Immunohistochemistry was performed with: anti-human fibroblast (CD90), anti-human epithelial antigen (Ber-EP4, CD49f), anti-human endometriotic stromal marker (CD10) and estrogen receptor-a (ER-a). Results: Two types of colonies were observed: small loosely packed colonies containing large cells and large colonies with small, densely packed cells. The total CE for epithelial endometriotic cells was 2.92+2.49% (n ¼ 19); 1.70+1.47% for large and 1.22+1.02% for small colonies. Whereas, the total CE for stromal endometriotic cells was 4.67+2.71% (n ¼ 19); 1.54+0.92% for large and 3.13+1.80% for small colonies. There was no significant difference between the two cell types. Single cells derived from large epithelial and stromal endometriotic clones could be subcloned four times more when compared to those from small clones (n ¼ 5). Small epithelial colonies were positive for epithelial marker and negative for the fibroblast marker. However, large colonies were negative for BerEP4 and CD49f suggesting these epithelial markers are lost during in vitro proliferation. Both large and small stromal colonies were positive for CD90 and CD10 and were negative for epithelial markers. Interestingly, both large epithelial and stromal endometriotic clones possessed ER-a immunoreactivity. Conclusions: These results suggest that a small population of cells with stemlike cell properties of clonogenic and self renewal activity may be responsible for the development and progression in endometriosis. References Wheeler, J.M., Epidemiology of endometriosis-associated infertility. J Reprod Med, 1989. 34(1): p. 41–6. Chan, R.W., K.E. Schwab, and C.E. Gargett, Clonogenicity of human endometrial epithelial and stromal cells. Biol Reprod, 2004. 70(6): p. 1738–50. Chan, R.W. and C.E. Gargett, Identification of label-retaining cells in mouse endometrium. Stem Cells, 2006. 24(6): p. 1529–38.-
dc.languageeng-
dc.publisherOxford University Press. The Journal's web site is located at http://humrep.oxfordjournals.org/-
dc.relation.ispartofHuman Reproduction-
dc.rightsPre-print: Journal Title] ©: [year] [owner as specified on the article] Published by Oxford University Press [on behalf of xxxxxx]. All rights reserved. Pre-print (Once an article is published, preprint notice should be amended to): This is an electronic version of an article published in [include the complete citation information for the final version of the Article as published in the print edition of the Journal.] Post-print: This is a pre-copy-editing, author-produced PDF of an article accepted for publication in [insert journal title] following peer review. The definitive publisher-authenticated version [insert complete citation information here] is available online at: xxxxxxx [insert URL that the author will receive upon publication here].-
dc.titleHuman endometriotic cells exhibit stem-like cell properties-
dc.typeConference_Paper-
dc.identifier.emailChan, RWS: rwschan@HKUCC-COM.hku.hk-
dc.identifier.emailNg, EHY: nghye@hkucc.hku.hk-
dc.identifier.emailLee, CKF: ckflee@hkucc.hku.hk-
dc.identifier.emailYeung, WSB: wsbyeung@hkucc.hku.hk-
dc.identifier.authorityNg, EHY=rp00426-
dc.identifier.authorityLee, CKF=rp00458-
dc.identifier.authorityYeung, WSB=rp00331-
dc.description.natureabstract-
dc.identifier.doi10.1093/humrep/dep759-
dc.identifier.hkuros160201-
dc.identifier.volume24-
dc.identifier.issueS1-
dc.identifier.spagei97-
dc.identifier.epagei97-
dc.publisher.placeUnited Kingdom-

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