Yardımcı üreme tekniklerinde endometriyal kokültür hücreleri

Amaç: Endometriyum, üreme biyolojisinde çok önemli bir rol oynar ve ona bağlı olarak gelişen patolojilerden biri implantasyonda yaşanan olumsuzluklardır. İmplantasyon başarısızlıkları, kadın infertilitesi için önemli süreçlerdendir ve çözümüne dair geliştirilen yöntemlerden biri ise endometriyal kokültür uygulamalarıdır. Bu çalışmada, in vitro monolayer endometriyal kokültürdeki hücre tiplerinin ayırt edilmesi, doku dinamiğinin aydınlatılabilmesi ve klinik uygulamalarda kullanımı açısından yararlılığının gösterilmesi amaçlandı. Yöntemler: Çalışmamıza 8 fertil ve Tekrarlayan İmplantasyon Başarısızlığı (TİB) yaşayan 16 infertil kadından oluşan 2 grup dâhil edildi. Negatif basınçlı pipelle aspirasyon yapılarak endometriyal doku örnekleri elde edildi. Embriyoyu blastosist aşamasına götüreceği düşünülen "Endometriyal Epitel-Gland hücresi (EE)" ve "Stromal (ES) hücreler" ayrıştırılıp kültürleri yapıldı. Daha sonra bu dokular ışık mikroskobik doku takibine alındı. Kokültür hücreleri, toluidin mavisi ile boyandıktan sonra gruplandırıldı. Hücre tiplerine ait sayısal değerler ile çalışmanın istatiksel analizi ki-kare metodu kullanılarak yapıldı. Bulgular: Kokültürlerde hem fertil hem de TİB grubundan elde edilen hücre tipleri ışık mikroskobik incelemeyle morfolojik özelliklerine göre ayırt edildi. Her iki grupta da, vakuollü, mikrovilluslu ve sitoplazmik uzantılı olarak isimlendirilen hücreler tanımlandı. Sonuç: Her iki grubun, endometriyum dokularından elde edilen kokültürleri ışık mikroskobik düzeyde incelendi. Böylece gruplar arası farklılıkların belirlenmesi kokültürdeki hücre yapılarının sınıflandırılması sağlandı ve endometriyal kokültür uygulamalarının embriyonun gelişiminde faydalı olabileceği kanısına varıldı.

Endometrial cocultured cells in assisted reproduction techniques

Objective: Endometrium, which plays a very important role in reproductive biology and one of the pathologies connected with it as are the problems in the implantation. One of the most important processes for the female fertility are experienced in implantation failure and one of the developed methods for the solution are endometrial coculture application. In this study, to provide knowledge of the types of cells in in vitro monolayer endometrial coculture. Methods: In our study, consisting of 8 fertile and 16 infertile women suffering from recurrent implantation failure were included in two groups. Performing aspiration with negative pressure pipeline endometrial tissue samples were obtained. Intended to carry embryo to the blastocyst stage, Endometrial Epithelial-gland cells (EG) and Epithelial stromal (ES) cells are obtained, after concluding of the culture was provided to do light microscopic tissue follow of these tissues. Cocultured cells were grouped stained with toluidine blue. Working with the numerical values of the cell types was performed using the Chi-square statistical analysis method.Results: In cocultured, both fertile and cell types derived from the TIB group, with light microscopic examination, were distinguished by morphological characteristics. Each of the two groups, called cells called vacuoles, microvilli and cytoplasmic extension was defined.Conclusion: Both groups, coculture derived from endometrial tissue were examined by light microscope. Thus, the identification of differences between groups was provided classification of cell structures in the coculture and endometrial cocultured practices, was concluded to be beneficial in the embryo's development.

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Maugey-Laulom B, Commenges-Ducos M, Jullien V, et al. Endometrial vascularity and on going preg- nanacy after IVF. Eur J Obstet Gynecol Reprod Bio 2002;104:137-143.
Bahar L, Baykal T. Endometriyal reseptivitenin im- plantasyondaki rolü. Mersin Univ Saglık Bilim Derg 2008;1:1-6.
Polanski LT, Baumgarten MN, Quenby S, et al. What exactly do we mean by 'recurrent implantation failure'? A systematic review and opinion. Reprod Biomed On- line 2014;28:409-423.
Singh M, Chaudhry P, Asselin E. Bridging endometrial receptivity and implantation: network of hormones, cy- tokines, and growth factors. J Endocrinol 2011;210:5- 14.
Bee K, Tan BK, Vandekerckhove P, et al. Investigation and current management of recurrent IVF treatment failure in the UK. Int J O&G 2005;112:773-780.
Coughlan C, Ledger W, Wang Q, et al. Recurrent im- plantation failure: definition and management. Reprod Biomed Online 2014;28:14-38.
Achache H, Revel A. Endometril receptivity markers, the journey to successful embryo implantation. Hum Reprod Update 2006;12:731-746.
Racicot KE, Wu¨nsche V, Auerbach B, et al. Human chorionic gonadotropin enhances trophoblast-epithe- lial ınteraction in an in vitro model of human implanta- tion. Reprod Sci 2014;21:1274-1280.
Nardo LG. Human embriyo implantation failure and recurrent miscarriage: basic science and clinical. Re- prod Biomed Online 2006;13:11-2.
Plachot M, Alvarez S, Merviel P, et al. Role of endo- metrial cells in "in vitro" embryo development. Assist Reprod Rev 1994;4:85-95.
Bahar L, Eras N. Endometrial ko-kültür uygulamaları. Genel Tıp Derg 2012;22:31-36.
Kim YB, Ahn SH, Chang DY, et al. Vero cell co-culture counteracts the detrimental effects of hydrosalpinx fluid on the development of mouse embryos in vitro. J Korean Med Sci 2002;17:217-219.
Nottola SA, Heyn R, Camboni A, et al. Ultrastructural characteristics of human granulosa cells in a cocul- ture system for in vitro fertilization. Microsc Res Tech 2006;69:508-516.
Bongso A, Fong, CY, Ng SC. Human embryonic be- havior in a sequencial human oviduct-endometrial co- culture system. Fertil Steril 1994;61:976-978.
Barmat LI, Liu H-C, Spandorfer CD, et al. Human preembryo development on autologous endometrial coculture versus conventional medium: A randomized trial. Fertil Steril 1998;70:1109-1113.
Barmat LI, Liu H-C, Spandorfer SD, et al. Autologous endometrial co-culture in patients with repeated fail- ures of implantation after in vitro fertilization-embryo transfer. J Assist Reprod Genet 1999;16:121-127.
Seferli R, Tokmak A, Ersoy AÖ, et al. Assessment of reproductive results of infertile patients who had un- dergone operative hysteroscopy for intrauterine pa- thology. J Clin Exp Invest 2015;6:52-58.
Ledee-Bataille N, Lapree-Delage G, Taupin JL. Con- centration of leukaemia inhibitory factor (LIF) in uter- ine flushing fluid is highly predictive of embryo implan- tation. Hum Reprod 2002;17:213-218.
Cavalcante MB, Costa FD, Barini R, Jşnior EA. Gran- ulocyte colony-stimulating factor and reproductive medicine: A review. Iran J Reprod Med 2015;13:195- 202.
Tada Y, Kitaya K, Amano N, et al. A pilot survey on obstetric complications in pregnant women with a his- tory of repeated embryo implantation failure and those undergoing single local endometrial injury. Clin Exp Obstet Gynecol 2015;42:176-178.
Potdar N, Gelbaya T, Nardo LG. Endometrial injury to overcome recurrent embryo implantation failure: a systematic review and meta-analysis. Reprod Biomed Online 2012;25:561-571.
Zhou L, Li R, Wang R, et al. Local injury to the en- dometrium in controlled ovarian hyperstimulation cycles improves implantation rates. Fertil Steril 2008;89:1166-1176.
Zhang XH, Liu ZZ, Tang MX, et al. Morphological changes and expression of cytokine after local endo- metrial injury in a mouse model. Reprod Sci 2015; doi: 10.1177/1933719115580999.
Melnick AP, Murphy EM, Masbou AK, et al. Autolo- gous endometrial coculture biopsy: is timing every- thing? Fertil Steril 2015;104:104-109.
Simón C, Mercader A, Garcia-Velasco J, et al. Cocul- ture of human embryos with autologous human endo- metrial human endometrial epithelial cells in patients with implantation failure. J Clin Endocrinol Metab 1999;84:2638-2646.
Urman B, Yakın K, Balaban B. Recurrent implanta- tion failure in assisted reproduction: how to counsel and manage. General considerations and treatment options that may benefit the couple. RBM Online 2005;11:371-381.
Spandorfer SD, Barmat LI, Liu H-C, et al. Granulo- cyte Macrophage-Colony Stimulating Factor produc- tion by autologous endometrial co-culture is associ- ated with outcome for IVF patients with a history of multiple implantation failures. Am J Reprod Immunol 1998;40:377-381.
Spandorfer SD, Soslow R, Clark R, et al. Histologic characteristics of the endometrium predicts suc- cess when utilizing autologous endometrial coculture in patients with IVF failure. J Assist Reprod Genet 2006;23:185-189.
Strowitzki T, Germeyer A, Popovici R, Wolff M. The human endometrium as a fertility-determining factor. Hum Reprod Update 2006;12:617-630.
Dimitriadis E, White CA, Jones RL, Salamonsen LA. Cytokines, chemokines and growth factors in endo- metrium related to implantation Hum Reprod Update 2005;11:613-630.
İskender B, İzgi K, Şanlıoğlu S, Canatan H. Human embryonic stem cells and microenvironment. J Clin Exp Invest 2014;5:486-495.