Isolation of human uterine smooth muscle cells     

1. Approval to utilize human tissue was given by Hospital's Committee on Research Involving Human Subjects.
2. Human myometrial and leiomyoma tissues were obtained from surplus pieces of surgical specimens after hysterectomies.
3. Matched myometrium and leiomyoma tissues were processed and cultured with Primary cell isolation kit and primary cell culture.
4. The cultures were never passaged and were used within 2 weeks of the initial plating.
5. The source of the tissues dictated and generally limited the number of culture dishes that could be prepared for analysis (generally 10-20 culture plates for one experiment).
6. Twenty-four hours prior to the experiments, the culture medium was changed to serum-free, primary cell medium supplemented.
7. All experiments with the primary cultures were performed using this defined serum-free medium.

References
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4. Berthois Y., Katzenellenbogen J. A., Katzenellenbogen B. S. (1986) Proc. Natl. Acad. Sci. U. S. A. 83, 2496–2500.