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验证激光捕获显微切割方案在子宫内膜异位症研究中的应用。

Validation of Laser Capture Microdissection Protocol in Endometriosis Studies.

机构信息

Hospital Israelita Albert Einstein, São Paulo, BR 05652-900, Brazil.

Centro de Endometriose São Paulo, São Paulo, BR 04534-001, Brazil.

出版信息

Medicina (Kaunas). 2019 Aug 22;55(9):520. doi: 10.3390/medicina55090520.

Abstract

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. 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. We obtained satisfactory results regarding the yields and integrity of RNA and protein obtained from LCM-processed endometriosis tissues. LCM can provide more precise analysis of endometriosis biopsies, provided that key steps of the methodology are followed.

摘要

子宫内膜样组织出现在子宫腔外是子宫内膜异位症的一个关键特征。尽管子宫内膜异位病灶在组织学上似乎与在位子宫内膜非常相似,但需要对两者进行详细比较研究,因为它们的内部和周围细胞排列是不同的。因此,组织之间的比较可能需要使用一些方法,如激光捕获显微切割(LCM),以实现对特定区域及其特定细胞群的精确选择。然而,众所周知,LCM 的有效使用取决于研究组织的类型和选择合适的方案。最近的研究已经报道了在子宫内膜异位症研究中使用 LCM。本研究的主要目的是建立一个标准化的方案,从在位或异位子宫内膜中获得高质量的微切割材料。为了进一步应用于分子技术,我们对 LCM 处理的病变样本的主要方法学步骤进行了标准化,以获得高质量的材料。我们从 LCM 处理的子宫内膜异位症组织中获得了满意的 RNA 和蛋白质产量和完整性结果。只要遵循该方法学的关键步骤,LCM 就可以提供更精确的子宫内膜异位症活检分析。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0b2c/6780092/12297258bbc8/medicina-55-00520-g001.jpg

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