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Published on: January 26, 2018
Development and validation of a three-dimensional in vitro model for uterine leiomyoma and patient-matched myometrium
Minnie Malik1, William H Catherino
1Department of Obstetrics and Gynecology, Uniformed Services University of the Health Sciences, Bethesda, Maryland 20814-4799, USA.
Objective:
To develop and validate a three-dimensional (3D) culture system of leiomyoma and myometrial cells.
Design:
In vitro study of immortalized cultures of patient-matched leiomyoma and myometrium.
Setting:
University hospital.
Patient(S):
Women undergoing hysterectomy for symptomatic leiomyomas.
Intervention(S):
Immortalized cell cultures, quantitative reverse-transcriptase polymerase chain reaction (qRT-PCR), cytoimmunofluorescence, and Western blot analysis.
Main Outcome Measure(S):
Morphologic features; expression of COL1A1, versican, fibronectin, dermatopontin, and transforming growth factor β3 (TGF-β3); and integrin-mediated 3D structural formation.
Result(S):
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.
Conclusion(S):
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.
