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Validation of Laser Capture Microdissection Protocol in Endometriosis Studies

Background and Objectives: The presence of endometrial-like tissue outside the uterine cavity is a key feature of endometriosis. Although endometriotic lesions appear to be histologically quite similar to the eutopic endometrium, detailed studies comparing both tissues are required because their inn...

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Autores principales: da Costa, Katiane de Almeida, Malvezzi, Helena, Viana, Bruno Gallani, Filippi, Renée Zon, Neme, Rosa Maria, Aloia, Thiago Pinheiro Arrais, Podgaec, Sérgio, Piccinato, Carla de Azevedo
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6780092/
https://www.ncbi.nlm.nih.gov/pubmed/31443551
http://dx.doi.org/10.3390/medicina55090520
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author da Costa, Katiane de Almeida
Malvezzi, Helena
Viana, Bruno Gallani
Filippi, Renée Zon
Neme, Rosa Maria
Aloia, Thiago Pinheiro Arrais
Podgaec, Sérgio
Piccinato, Carla de Azevedo
author_facet da Costa, Katiane de Almeida
Malvezzi, Helena
Viana, Bruno Gallani
Filippi, Renée Zon
Neme, Rosa Maria
Aloia, Thiago Pinheiro Arrais
Podgaec, Sérgio
Piccinato, Carla de Azevedo
author_sort da Costa, Katiane de Almeida
collection PubMed
description Background and Objectives: The presence of endometrial-like tissue outside the uterine cavity is a key feature of endometriosis. Although endometriotic lesions appear to be histologically quite similar to the eutopic endometrium, detailed studies comparing both tissues are required because their inner and surrounding cellular arrangement is distinct. Thus, comparison between tissues might require methods, such as laser capture microdissection (LCM), that allow for precise selection of an area and its specific cell populations. However, it is known that the efficient use of LCM depends on the type of studied tissue and on the choice of an adequate protocol. Recent studies have reported the use of LCM in endometriosis studies. The main objective of the present study is to establish a standardized protocol to obtain good-quality microdissected material from eutopic or ectopic endometrium. Materials and Methods: The main methodological steps involved in the processing of the lesion samples for LCM were standardized to yield material of good quality to be further used in molecular techniques. Results: We obtained satisfactory results regarding the yields and integrity of RNA and protein obtained from LCM-processed endometriosis tissues. Conclusion: LCM can provide more precise analysis of endometriosis biopsies, provided that key steps of the methodology are followed.
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spelling pubmed-67800922019-10-30 Validation of Laser Capture Microdissection Protocol in Endometriosis Studies da Costa, Katiane de Almeida Malvezzi, Helena Viana, Bruno Gallani Filippi, Renée Zon Neme, Rosa Maria Aloia, Thiago Pinheiro Arrais Podgaec, Sérgio Piccinato, Carla de Azevedo Medicina (Kaunas) Article Background and Objectives: The presence of endometrial-like tissue outside the uterine cavity is a key feature of endometriosis. Although endometriotic lesions appear to be histologically quite similar to the eutopic endometrium, detailed studies comparing both tissues are required because their inner and surrounding cellular arrangement is distinct. Thus, comparison between tissues might require methods, such as laser capture microdissection (LCM), that allow for precise selection of an area and its specific cell populations. However, it is known that the efficient use of LCM depends on the type of studied tissue and on the choice of an adequate protocol. Recent studies have reported the use of LCM in endometriosis studies. The main objective of the present study is to establish a standardized protocol to obtain good-quality microdissected material from eutopic or ectopic endometrium. Materials and Methods: The main methodological steps involved in the processing of the lesion samples for LCM were standardized to yield material of good quality to be further used in molecular techniques. Results: We obtained satisfactory results regarding the yields and integrity of RNA and protein obtained from LCM-processed endometriosis tissues. Conclusion: LCM can provide more precise analysis of endometriosis biopsies, provided that key steps of the methodology are followed. MDPI 2019-08-22 /pmc/articles/PMC6780092/ /pubmed/31443551 http://dx.doi.org/10.3390/medicina55090520 Text en © 2019 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
da Costa, Katiane de Almeida
Malvezzi, Helena
Viana, Bruno Gallani
Filippi, Renée Zon
Neme, Rosa Maria
Aloia, Thiago Pinheiro Arrais
Podgaec, Sérgio
Piccinato, Carla de Azevedo
Validation of Laser Capture Microdissection Protocol in Endometriosis Studies
title Validation of Laser Capture Microdissection Protocol in Endometriosis Studies
title_full Validation of Laser Capture Microdissection Protocol in Endometriosis Studies
title_fullStr Validation of Laser Capture Microdissection Protocol in Endometriosis Studies
title_full_unstemmed Validation of Laser Capture Microdissection Protocol in Endometriosis Studies
title_short Validation of Laser Capture Microdissection Protocol in Endometriosis Studies
title_sort validation of laser capture microdissection protocol in endometriosis studies
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6780092/
https://www.ncbi.nlm.nih.gov/pubmed/31443551
http://dx.doi.org/10.3390/medicina55090520
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