title: Optimized culture conditions for tissue explants of uterine leiomyoma.
abstract: BACKGROUND
  Uterine leiomyomas are the most common benign tumours in women, which arise from smooth muscle 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 culturing 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 proliferation and apoptotic indices and real-time PCR were performed.
  
  
  RESULTS
  Main parameters for culturing myoma tissue explants were tested for finding an optimal protocol. Standard 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 established tissue culture model allows examining the effects of known and potential therapeutic substances and the influence of immune competent cells in the process of tumour formation to find new targets for medical treatment.
authors.affiliation: Department of Gynaecology, Charité Hospital Campus Benjamin Franklin, Berlin, Germany.
authors.forename: A
authors.initials: A
authors.lastname: Fiebitz
authors.forename: M
authors.initials: M
authors.lastname: Fritsch
authors.forename: U
authors.initials: U
authors.lastname: Reichelt
authors.forename: C
authors.initials: C
authors.lastname: Ruester
authors.forename: V
authors.initials: V
authors.lastname: Chiantera
authors.forename: G F
authors.initials: GF
authors.lastname: Vercellino
authors.forename: A
authors.initials: A
authors.lastname: Darwish
authors.forename: A
authors.initials: A
authors.lastname: Schneider
authors.forename: S
authors.initials: S
authors.lastname: Mechsner
mesh_terms: D003470:Culture Media; D004815:Epidermal Growth Factor / Q000008:administration & dosage; D004958:Estradiol / Q000008:administration & dosage; D005260:Female; D006801:Humans; D007150:Immunohistochemistry; D007889:Leiomyoma / Q000473:pathology*; D011374:Progesterone / Q000008:administration & dosage; D012333:RNA, Messenger / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D014594:Uterine Neoplasms / Q000473:pathology*
chemical_list: D003470:Culture Media; D012333:RNA, Messenger; D011374:Progesterone; D004958:Estradiol; D004815:Epidermal Growth Factor