Use of laser capture microdissection in studying hormone-dependent diseases: endometriosis

Sachiko Matsuzaki, Michel Canis, Gérard Mage
Methods in molecular biology (Clifton, N.J.) · 2009 · doi:10.1007/978-1-60327-378-7_19 · PMID:19763512
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⚙ AI-generated summary by claude@2026-06, 2026-06-10 ⓘ

Laser capture microdissection provides pure cell populations from endometrial and endometriotic tissues, enabling high-quality RNA extraction for differential gene expression analysis in hormone-dependent diseases.

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This protocol chapter describes how to use laser capture microdissection to quantify gene expression separately in specific cell populations within heterogeneous endometriotic tissue, rather than relying on bulk homogenates that reflect mixed cell types (including epithelial, stromal, fibrotic, and muscle components). It focuses on obtaining high-quality RNA from frozen endometrial and endometriotic tissues for downstream applications, using the Arcturus PixCell II system. A key caveat emphasized by the broader context of such work is that RNA integrity/quality can affect gene expression measurements, motivating careful RNA handling. Relevance to endometriosis: the paper is an endometriosis-focused methods chapter that frames laser capture microdissection as essential for identifying genetic markers by separating endometriotic cell populations, addressing endometriosis-specific heterogeneity.

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Abstract

Endometriosis, a common gynecological disorder responsible for infertility and pelvic pain, is defined as the presence of endometrial glands and stroma within extra-uterine sites. Gene expression studies performed on endometriotic tissue homogenates have yielded results reflecting mRNA abundance in a mixture of cell types (including epithelial cells, stromal cells, fibrotic tissue, and muscle tissue). Therefore, a method for quantifying gene expression separately in individual cell populations is essential for identifying genetic markers. Laser capture microdissection is a technique for obtaining pure populations of cells from heterogeneous tissues. This chapter provides methods to obtain high-quality RNA suitable for a variety of different down stream applications from frozen endometrial and endometriotic tissues for laser capture microdissection, using the Arcturus PixCell II system.
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Abstract

Endometriosis, a common gynecological disorder responsible for infertility and pelvic pain, is defined as the presence of endometrial glands and stroma within extra-uterine sites. Gene expression studies performed on endometriotic tissue homogenates have yielded results reflecting mRNA abundance in a mixture of cell types (including epithelial cells, stromal cells, fibrotic tissue, and muscle tissue). Therefore, a method for quantifying gene expression separately in individual cell populations is essential for identifying genetic markers. Laser capture microdissection is a technique for obtaining pure populations of cells from heterogeneous tissues. This chapter provides methods to obtain high-quality RNA suitable for a variety of different down stream applications from frozen endometrial and endometriotic tissues for laser capture microdissection, using the Arcturus PixCell II system. Access this chapter Tax calculation will be finalised at checkout Purchases are for personal use only Similar content being viewed by others

References

Matsuzaki S, Canis M, Vaurs-Barriere C, Pouly JL, Boespflug-Tanguy O, Penault-Llorca F, Dechelotte P, Dastugue B, Okamura K, Mage G. (2004) DNA microarray analysis of gene expression profiles in deep endometriosis using laser capture microdissection. Mol Hum Reprod. 10:719–28. Matsuzaki S, Canis M, Vaurs-Barriere C, Boespflug-Tanguy O, Dastugue B, Mage G. (2005) DNA microarray analysis of gene expression in eutopic endometrium from patients with deep endometriosis using laser capture microdissection. Fertil Steril, 84 Suppl 2:1180–90. Emmert-Buck, M.R., Bonner, R.F., Smith, P.D., Chuaqui, R.F., Zhuang, Z., Goldstein, S.R., Weiss, R.A. and Liotta, L.A. (1996) Laser capture microdissection. Science, 274, 998–1001. Fleige S, Pfaffl MW. (2006) RNA integrity and the effect on the real-time qRT-PCR performance. Mol Aspects Med. 2006; 27:126–39. Fleige S, Walf V, Huch S, Prgomet C, Sehm J, Pfaffl MW (2006). Comparison of relative mRNA quantification models and the impact of RNA integrity in quantitative real-time RT-PCR. Biotechnol Lett. 28:1601–13. Schroeder A, Mueller O, Stocker S, Salowsky R, Leiber M, Gassmann M, Lightfoot S, Menzel W, Granzow M, Ragg T (2006). The RIN: an RNA integrity number for assigning integrity values to RNA measurements. BMC Mol Biol. 7:3. Strand C, Enell J, Hedenfalk I, Fernö M (2007). RNA quality in frozen breast cancer samples and the influence on gene expression analysis––a comparison of three evaluation methods using microcapillary electrophoresis traces. BMC Mol Biol. 8:38. Gassmann, M. (2003) Quality Assurance of RNA derived from laser microdissected tissue samples obtained by the PALM(R) MicroBeam System using the RNA 6000 Pico LabChip(R) kit. 1-8. Agilent Technologies. Acknowledgments This work was supported in part by Karl Storz Endoscopy & GmbH (Tuttlingen, Germany) and le Conseil Régional d’Auvergne (Recherche et Innovation Technologique) (Clermont-Ferrand, France). Author information Authors and Affiliations Editor information Editors and Affiliations Rights and permissions Copyright information © 2009 Humana Press, a part of Springer Science+Business Media, LLC About this protocol Cite this protocol Matsuzaki, S., Canis, M., Mage, G. (2009). Use of Laser Capture Microdissection in Studying Hormone-Dependent Diseases: Endometriosis. In: Park-Sarge, OK., Curry, T. (eds) Molecular Endocrinology. Methods in Molecular Biology, vol 590. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60327-378-7_19 Download citation DOI: https://doi.org/10.1007/978-1-60327-378-7_19 Published: Publisher Name: Humana Press, Totowa, NJ Print ISBN: 978-1-60327-377-0 Online ISBN: 978-1-60327-378-7 eBook Packages: Springer Protocols

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Condition tags

endometriosisinfertility

MeSH descriptors

Endometriosis Gonadal Steroid Hormones Biopsy Endometriosis Endometriosis Female Gonadal Steroid Hormones Humans

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