0.4 mm porous membranes

Human primary or immortalized myometrium and leiomyoma cells were cocultured with human adipocyte (SW872) cells in a Transwell system using 0.4-mm porous membranes from Corning (Corning, New York, USA) with their respective media. We observed a non-significant inter-individual variance in adipocyte responses in primary myometrium and leiomyoma cells from different subjects. We have not pooled cells from different patients together. Similarly, rat leiomyoma cells (ELT3) and differentiated mouse 3T3-L1 cells were cocultured in a Transwell system. The Transwell system was cultured for eight days, with 50% of the medium being changed every 48 h. In other sets of experiments, the cultured (single cell type) and cocultured (with adipocytes) cells were treated with a TNF-α inhibitor (SPD-304), an MCP-1 inhibitor (bindarit), and a nuclear factor kappa B (NF-κB) inhibitor (withaferin A) on day 7 then left for 24 h. After the incubation, the myometrium and leiomyoma media were collected for the multiplex array, and the cells were harvested for protein quantification and analysis.
The use of coculture systems may better elucidate the interaction between myometrium/leiomyoma cells and adipocytes. This system also allows us to examine the aggregated impact on the cellular secretome (Fig. 1).

Human primary or immortalized myometrium and leiomyoma cells were cocultured with human adipocyte (SW872) cells in a Transwell system using 0.4-mm porous membranes from Corning (Corning, New York, USA) with their respective media. We observed a non-significant inter-individual variance in adipocyte responses in primary myometrium and leiomyoma cells from different subjects. We have not pooled cells from different patients together. Similarly, rat leiomyoma cells (ELT3) and differentiated mouse 3T3-L1 cells were cocultured in a Transwell system. The Transwell system was cultured for eight days, with 50% of the medium being changed every 48 h. In other sets of experiments, the cultured (single cell type) and cocultured (with adipocytes) cells were treated with a TNF-α inhibitor (SPD-304), an MCP-1 inhibitor (bindarit), and a nuclear factor kappa B (NF-κB) inhibitor (withaferin A) on day 7 then left for 24 h. After the incubation, the myometrium and leiomyoma media were collected for the multiplex array, and the cells were harvested for protein quantification and analysis.
The use of coculture systems may better elucidate the interaction between myometrium/leiomyoma cells and adipocytes. This system also allows us to examine the aggregated impact on the cellular secretome (Fig. 1).