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Abstract

Optimized Culture Conditions for Tissue Explants of Uterine Leiomyoma by A. Fiebitz, M. Fritsch, U. Reichelt, C. Ruester, v. Chiantera, G. F. Vercellino, A. Darwish, A. Schneider, S. Mechsner

Background: Uterine leiomyomas are the most common benign tumours in women, which arise from smooth mus-cle cells of the uterine myometrium and usually are multicentric. In spite of their frequency pathogenesis is widely unknown, mainly due to the absence of a suitable model system. We describe the systematic optimization of cul-turing leiomyoma tissue explants in an economical and effective ex vivo system.
Methods: Different concentrations of oxygen, different media, sera, hormones, and growth factor supplements were tested. Immunohistochemical stainings with antibodies against hormone receptors as well as specifying pro-liferation and apoptotic indices and real-time PCR were performed.
Results: Main parameters for culturing myoma tissue explants were tested for finding an optimal protocol. Stan-dard medium D-MEM-F12 in combination with the use of horse serum in a reduced concentration of 1% turned out to be optimal for these tissue cultures as well as the addition of estradiol and epidermal growth factor EGF to media. Reduced oxygen content in the incubator air showed no positive effect.
Conclusions: For culturing tissue explants of uterine leiomyoma several conditions were optimized. The estab-lished tissue culture model allows examining the effects of known and potential therapeutic substances and the in-fluence of immune competent cells in the process of tumour formation to find new targets for medical treatment.

DOI: 10.7754/Clin.Lab.2012.111117