Abstract
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.
Highlights
Endometriosis is a disease characterized by the presence of endometrial-like tissue outside the uterine cavity [1]
Because of the morphological similarity between endometriotic lesions and normal endometrium, researchers have investigated both tissues to find characteristic genetic or protein markers related to the pathophysiology of endometriosis
We have shown in the present work, for the first time, encouraging results on the recovery of RNA and protein from endometrium and endometriotic lesion tissue sections processed by laser capture microdissection (LCM)
Summary
Endometriosis is a disease characterized by the presence of endometrial-like tissue outside the uterine cavity [1]. The stroma of endometriotic lesions is surrounded and infiltrated by immune, inflammatory and vascular system cells in numbers and types that are distinct from those found in the eutopic endometrium [3]. Considering that distinct cell types express various genes and proteins at different levels, comparisons between tissue samples with diverse cellular infiltrates would fail to accurately answer specific quests on gene and protein expression, especially when only a minimum amount of material is available for analysis. 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. 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. Conclusion: LCM can provide more precise analysis of endometriosis biopsies, provided that key steps of the methodology are followed
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