Establishment of a 3D spheroid culture system to evaluate the responsiveness of uterine leiomyoma cells to female hormones

Abstract

Abstract Purpose Uterine leiomyomas (ULMs) are classified into those with and without MED12 mutations (MED12m(+) and MED12m(−), respectively). This study was undertaken to establish a culture system to evaluate the effect of female hormones on the growth of ULM cells in each ULM subtype. Methods ULM cells isolated from MED12m(+) or MED12m(−) tissues were cultured in a monolayer for 7 days with four hormone treatments: estrogen (E) and progesterone (P) (E + P), E only (E), P only (P), and medium only (CTRL). They were also cultured in a 3D spheroid culture system with the above four treatments and a fifth treatment: E + P + selective progesterone receptor modulator (E + P + SPRM). The hormonal effects were evaluated based on cell number, spheroid size, and histology. Results In the monolayer cultures, female hormones did not cause the proliferation of ULM cells of either subtype. In the spheroid cultures, spheroid sizes for both subtypes were significantly larger with the E + P and P treatments than with the CTRL and E treatments and were comparable in the E and E + P + SPRM treatments. Histological staining showed that collagen fibers were present only in the spheroids of the P‐treated groups of MED12m(+). Conclusion We established a 3D spheroid culture system to evaluate the effects of female hormones on ULM cells.

Keywords

• female hormones
• mediator complex subunit 12 (MED12) mutation
• selective progesterone receptor modulator
• spheroid
• uterine leiomyomas