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

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.