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Validation of Laser Capture Microdissection Protocol in Endometriosis Studies.

Katiane de Almeida da CostaHelena MalvezziBruno Gallani VianaRenée Zon FilippiRosa Maria NemeThiago Pinheiro Arrais AloiaSérgio PodgaecCarla de Azevedo Piccinato
Published in: Medicina (Kaunas, Lithuania) (2019)
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.
Keyphrases
  • case control
  • gene expression
  • machine learning
  • stem cells
  • single cell
  • high speed
  • mass spectrometry
  • protein protein