Angiogenic properties of endometrial mesenchymal stromal cells in endothelial co-culture: an in vitro model of endometriosis.
Can endometrial mesenchymal stromal cells (E-MSCs) differentiate into endothelial cells in an in vitro co-culture system with human umbilical vein endothelial cells (HUVECs)?



E-MSCs can acquire endothelial markers and function in a direct co-culture system with HUVECs.



E-MSCs have been identified in the human endometrium as well as in endometriotic lesions. E-MSCs appear to be involved in formation of the endometrial stromal vascular tissue and the support of tissue growth and vascularization. The use of anti-angiogenic drugs appears as a possible therapeutic strategy against endometriosis.



This is an in vitro study comprising patients receiving surgical treatment of ovarian endometriosis (n = 9).



E-MSCs were isolated from eutopic and ectopic endometrial tissue and were characterized for the expression of mesenchymal and endothelial markers by FACS analysis and Real-Time PCR. CD31 acquisition was evaluated by FACS analysis and immunofluorescence after a 48 h-direct co-culture with green fluorescent protein +-HUVECs. A tube-forming assay was set up in order to analyze the functional potential of their interaction. Finally, co-cultures were treated with the anti-angiogenic agent Cabergoline.



A subpopulation of E-MSCs acquired CD31 expression and integrated into tube-like structures when directly in contact with HUVECs, as observed by both FACS analysis and immunofluorescence. The isolation of CD31+ E-MSCs revealed significant increases in CD31, vascular endothelial growth factor receptor 2, TEK receptor tyrosine kinase and vascular endothelial-Cadherin mRNA expression levels with respect to basal and to CD31neg cells (P < 0.05). On the other hand, the expression of mesenchymal genes such as c-Myc, Vimentin, neuronal-Cadherin and sushi domain containing 2 remained unchanged. Cabergoline treatment induced a significant reduction of the E-MSC angiogenic potential (P < 0.05 versus control).



Not applicable.



Further studies are necessary to investigate the cellular and molecular mechanisms underlying the endothelial cell differentiation.



E-MSCs may undergo endothelial differentiation, and be potentially involved in the development of endometriotic implants. Cell culture systems that more closely mimic the cellular complexity typical of endometriotic tissues in vivo are required to develop novel strategies for treatment.



This study was supported by the 'Research Fund ex-60%', University of Turin, Turin, Italy. All authors declare that their participation in the study did not involve actual or potential conflicts of interests.
Gynecology and Obstetrics I, Department of Surgical Sciences, Physiopathology of Reproduction and IVF Unit, S. Anna Hospital, Torino, Italy.
S
S
Canosa
Department of Molecular Biotechnology and Health Sciences, University of Torino, Torino, Italy.
A
A
Moggio
Department of Molecular Biotechnology and Health Sciences, University of Torino, Torino, Italy.
A
A
Brossa
Gynecology and Obstetrics I, Department of Surgical Sciences, Physiopathology of Reproduction and IVF Unit, S. Anna Hospital, Torino, Italy.
G
G
Pittatore
Gynecology and Obstetrics I, Department of Surgical Sciences, Physiopathology of Reproduction and IVF Unit, S. Anna Hospital, Torino, Italy.
G L
GL
Marchino
Gynecology and Obstetrics I, Department of Surgical Sciences, Physiopathology of Reproduction and IVF Unit, S. Anna Hospital, Torino, Italy.
S
S
Leoncini
Gynecology and Obstetrics I, Department of Surgical Sciences, Physiopathology of Reproduction and IVF Unit, S. Anna Hospital, Torino, Italy.
C
C
Benedetto
Gynecology and Obstetrics I, Department of Surgical Sciences, Physiopathology of Reproduction and IVF Unit, S. Anna Hospital, Torino, Italy.
A
A
Revelli
Department of Molecular Biotechnology and Health Sciences, University of Torino, Torino, Italy.
B
B
Bussolati
D020533:Angiogenesis Inhibitors / Q000494:pharmacology; D015415:Biomarkers / Q000378:metabolism; D000077465:Cabergoline; D015820:Cadherins / Q000235:genetics / Q000378:metabolism; D002450:Cell Communication / Q000187:drug effects; D002454:Cell Differentiation / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D018920:Coculture Techniques; D004715:Endometriosis / Q000235:genetics / Q000378:metabolism / Q000473:pathology* / Q000601:surgery; D004717:Endometrium / Q000378:metabolism / Q000473:pathology / Q000601:surgery; D004873:Ergolines / Q000494:pharmacology; D005260:Female; D015870:Gene Expression; D061307:Human Umbilical Vein Endothelial Cells / Q000166:cytology / Q000378:metabolism; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000187:drug effects / Q000378:metabolism / Q000473:pathology*; D008954:Models, Biological; D009389:Neovascularization, Pathologic / Q000235:genetics / Q000378:metabolism / Q000473:pathology*; D019408:Platelet Endothelial Cell Adhesion Molecule-1 / Q000235:genetics / Q000378:metabolism; D061251:Primary Cell Culture; D016271:Proto-Oncogene Proteins c-myc / Q000235:genetics / Q000378:metabolism; D042787:Receptor, TIE-2 / Q000235:genetics / Q000378:metabolism; D053669:Syndecan-2 / Q000235:genetics / Q000378:metabolism; D040301:Vascular Endothelial Growth Factor Receptor-2 / Q000235:genetics / Q000378:metabolism; D014746:Vimentin / Q000235:genetics / Q000378:metabolism
D020533:Angiogenesis Inhibitors; D015415:Biomarkers; D015820:Cadherins; D004873:Ergolines; C489427:MYC protein, human; D019408:Platelet Endothelial Cell Adhesion Molecule-1; D016271:Proto-Oncogene Proteins c-myc; C508245:SDC2 protein, human; D014746:Vimentin; D053669:Syndecan-2; C000588866:KDR protein, human; D042787:Receptor, TIE-2; D040301:Vascular Endothelial Growth Factor Receptor-2; D000077465:Cabergoline
CD31; anti-angiogenic therapy; cabergoline; co-culture; endometriosis; endothelial differentiation; mesenchymal stem cells