Stem Cell Growth and Differentiation in Organ Culture: New Insights for Uterine Fibroid Treatment

Organ culture allows for the understanding of normal and tumor cell biology, and tissues generally remain viable for 5-7 days. Strikingly, we determined that myometrial and MED12 mutant leiomyoma cells repopulated cell-depleted tissue slices after 20 days of culture. Using immunofluorescence and quantitative PCR of stem cell and undifferentiated cell markers, we observed clusters of CD49b(+) cells in tumor slices. CD49b(+) cells, however, were sparsely detected in the myometrial slices. Almost all LM cells strongly expressed Ki67, while only a few myometrial cells were stained for this proliferation marker. The CD73 marker was expressed only in tumor cells, whereas the mesenchymal stem cell receptor KIT was detected only in normal cells. HMGA2 and CD24 showed broader expression patterns and higher signal intensity in leiomyoma than in myometrial cells. In this study, we propose that activating CD49b(+) stem cells in myometrium leads to asymmetrical division, giving rise to transit-amplifying KIT+ cells that differentiate to smooth muscle cells. On the contrary, activated leiomyoma CD49b(+) cells symmetrically divide to form clusters of stem cells that divide and differentiate to smooth muscle cells without losing proliferation ability. In conclusion, normal and mutant stem cells can proliferate and differentiate in long-term organ culture, constituting a helpful platform for novel therapeutic discovery.

Automated summary

This study found that leiomyoma cells can repopulate cell-depleted tissue slices in organ culture, and clusters of CD49b(+) cells were observed in tumor slices. In contrast, CD49b(+) cells were sparsely detected in myometrial slices. Furthermore, leiomyoma cells strongly expressed Ki67 while myometrial cells only had a few stained cells. Based on these findings, it is proposed that activating CD49b(+) stem cells in the myometrium leads to asymmetrical division, giving rise to transit-amplifying KIT+ cells that differentiate to smooth muscle cells. On the other hand, activated leiomyoma CD49b(+) cells symmetrically divide to form clusters of stem cells that divide and differentiate to smooth muscle cells without losing proliferation ability.