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    Women's Health

    (M-489) A Method for Testing Compressive Forces on Fibroid Mimetic Cultures

    Thursday, October 12, 2023
    9:30 AM – 10:30 AM PDT
    Location: Exhibit Hall - Row M - Poster # 489
    Introduction:: Uterine fibroids are the most common benign tumors of the uterus affecting 1 in every 4 women and accounting for 40-60% of all hysterectomies. Common symptoms include heavy menstrual bleeding, pain, and infertility. Recent studies have investigated the effect of mechanical compression on uterine fibroids; however, most use a cell monolayer in which the cells do not produce significant extracellular matrix proteins and growth cannot be studied. Having a 3D model allows for more physiologically relevant results. The purpose of this study was to develop a model for the testing of compressive forces on tissue-engineered mimetic uterine fibroids.

    Materials and Methods:: Human myometrial and fibroid smooth muscle cells were isolated from biopsies obtained from patients undergoing myomectomy or hysterectomy for uterine fibroid treatment. All subjects provided informed consent. The spheroid model was developed by first centrifuging 200,000 cells in a microcentrifuge tube to form a cell pellet which was cultured for 5 days to promote adhesion. The spheroids were then embedded in 1% agarose gel in a 6-well plate with transwell inserts (1 mL of agarose for each insert). Compressive stress was applied by adding 15 g of weight (706.83 Pa) on top of the layer of agarose gel. Spheroid growth was measured by the area of the spheroids before and after adding the weights for 24 hours. Area was determined by image analysis using FIJI software. Statistical analysis was performed using a paired t-test with a 95% confidence interval.

    Results, Conclusions, and Discussions:: Preliminary results showed changes in size and appearance of the spheroids in culture; after 5 days there was a significant difference in growth of both fibroid and myometrial cell pellets (p< 0.05). However, these measurements were completed in collagen gels where cell migration made interpretations challenging. Hence the need to switch to an agarose substrate. After 24 hours of compression in agarose, spheroid area increased on average by approximately 0.191 mm2. However, the mean area of the spheroids after compression compared to before was found to not be significantly different (p=0.179).
    We have developed a fibroid mimetic culture that allows for the application of compressive forces to multiple spheroids at a time while still providing nutrients to the tissue and resulting in tissues where growth can be evaluated. While short-term compression for 24 hours showed no significant findings, preliminary data suggests that a minimum of 7 days in culture is necessary to determine differences in spheroid growth rates. This method will allow for the study of tissue growth and comparisons between different cell types, stimulation, or inhibitors to elucidate the mechanisms behind fibroid development and growth.


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