title: Development and validation of a three-dimensional in vitro model for uterine leiomyoma and patient-matched myometrium.
abstract: OBJECTIVE
  To develop and validate a three-dimensional (3D) culture system of leiomyoma and myometrial cells.
  
  
  METHODS
  In vitro study of immortalized cultures of patient-matched leiomyoma and myometrium.
  
  
  METHODS
  University hospital.
  
  
  METHODS
  Women undergoing hysterectomy for symptomatic leiomyomas.
  
  
  METHODS
  Immortalized cell cultures, quantitative reverse-transcriptase polymerase chain reaction (qRT-PCR), cytoimmunofluorescence, and Western blot analysis.
  
  
  METHODS
  Morphologic features; expression of COL1A1, versican, fibronectin, dermatopontin, and transforming growth factor β3 (TGF-β3); and integrin-mediated 3D structural formation.
  
  
  RESULTS
  Cells in 3D culture maintained spindle morphology. There was elevated expression of collagen 1A1 (6.66 ± 1.5), total versican (4.78 ± 0.5), fibronectin (3.94 ± 0.3), and TGF-β3 (2.21 ± 0.1) as was seen in progenitor tissue. Dermatopontin gene was down-regulated (0.29 ± 0.1), also similar to values in the surgical tissue sample. Myometrial cells in 3D culture responded to TGF-β3 with increased gene expression of collagen 1A1, fibronectin, and versican, and decreased expression of dermatopontin gene recapitulating the leiomyoma phenotype. Integrin-β1-inhibiting antibody disrupted the cell-extracellular matrix (ECM) communication and induced apoptosis.
  
  
  CONCLUSIONS
  Three-dimensional 3D leiomyoma cell cultures maintain the molecular phenotype of progenitor tissue, produce ECM, and interact with the ECM directly. This model system allows for assessment of the mechanism of aberrant ECM formation as well as the effectiveness of various potential therapies.
authors.affiliation: Department of Obstetrics and Gynecology, Uniformed Services University of the Health Sciences, Bethesda, Maryland 20814-4799, USA.
authors.forename: Minnie
authors.initials: M
authors.lastname: Malik
authors.forename: William H
authors.initials: WH
authors.lastname: Catherino
mesh_terms: D015153:Blotting, Western; D018929:Cell Culture Techniques / Q000379:methods* / Q000592:standards; D002454:Cell Differentiation / Q000502:physiology; D002461:Cell Line, Transformed; D011508:Chondroitin Sulfate Proteoglycans / Q000235:genetics / Q000378:metabolism; D024042:Collagen Type I / Q000235:genetics / Q000378:metabolism; D000090602:Collagen Type I, alpha 1 Chain; D016326:Extracellular Matrix Proteins / Q000235:genetics / Q000378:metabolism; D005260:Female; D005353:Fibronectins / Q000235:genetics / Q000378:metabolism; D015200:Fluoroimmunoassay; D006801:Humans; D007889:Leiomyoma / Q000473:pathology* / Q000503:physiopathology; D009215:Myometrium / Q000166:cytology* / Q000502:physiology; D061251:Primary Cell Culture; D020133:Reverse Transcriptase Polymerase Chain Reaction; D053782:Transforming Growth Factor beta3 / Q000235:genetics / Q000378:metabolism; D014594:Uterine Neoplasms / Q000473:pathology* / Q000503:physiopathology; D053675:Versicans / Q000235:genetics / Q000378:metabolism
chemical_list: D011508:Chondroitin Sulfate Proteoglycans; D024042:Collagen Type I; D000090602:Collagen Type I, alpha 1 Chain; C082019:DPT protein, human; D016326:Extracellular Matrix Proteins; D005353:Fibronectins; D053782:Transforming Growth Factor beta3; C508322:VCAN protein, human; D053675:Versicans