Comparative Studies on the In Vitro: MATERIALS AND METHODS(2)
Midluteal phase (9-10 days postovulation) endometrial tissue was obtained from adult female baboons by endo-metrectomy or after hysterectomy. Human endometrial tissue was obtained from endometrial biopsies or at hysterectomy from premenopausal women with no clinically documented abnormalities of the endometrium. All animal studies and the experimental use of human tissues were approved by the Animal Care Committee and the Institutional Review Board at the University of Illinois at Chicago, respectively. flovent inhaler
The tissue was minced thoroughly in calcium- and magnesium-free Hanks’ Balanced Salt Solution (HBSS). The minced tissue was placed in an enzyme solution containing 0.5% collagenase and 0.02% DNase and incubated for 20 min at 37°C in a shaking water bath. The supernatant was recovered and placed at 4°C. The remaining tissue was further digested in an enzyme solution consisting of 0.5% collagenase, 0.02% DNase, 0.1% hyaluronidase, and 0.1% pronase and processed as described above. The cell suspensions from the first and second digestions were centrifuged at 1500 X g for 5 min, and the pellet was resuspended in HBSS. The suspensions were then passed through a 95-^m nylon mesh followed by a 20-^m nylon mesh (Small Parts Inc., Miami, FL). The filtrates were pooled and centrifuged again at 1500 X g for 5 min.