Collection of CD-1 amniotic sacs
This protocol is extracted from research article:
Reversible EMT and MET mediate amnion remodeling during pregnancy and labor
Sci Signal, Feb 11, 2020; DOI: 10.1126/scisignal.aay1486

The Institutional Animal Care and Use Committee (IACUC) at the UTMB at Galveston approved the study protocol. CD-1 pregnant mice were purchased from Charles River Laboratories (Wilmington, MA). Animals were shipped on D10 of gestation and acclimated in a temperature- and humidity-controlled facility with automatically controlled 12-hour light/12-hour dark cycles. Mice were allowed to consume regular chow and drinking solution ad libitum. At day 14 of pregnancy, the pregnant CD-1 mice were weighed and subjected to minilaparotomy and injection of 150 μl of the treatment solution into uteri, physically between two to three gestational sacs, according to the following experimental groups: (i) CSE diluted in saline, (ii) CSE in combination with SB203580 (p38MAPK inhibitor), and (iii) saline alone (control). After euthanizing the animals by using carbon dioxide inhalation according to the IACUC and American Veterinary Medical Association guidelines on D18, maternal, fetal, and placental weight was documented, and pup loss/reabsorption was counted. Amniotic sacs and placentae were collected in either 10% formalin or fresh-frozen in liquid nitrogen and stored at −80°C until further analysis. In addition, CD-1 mice at embryonic D18 and D19 of pregnancy were euthanized using carbon dioxide inhalation according to the IACUC and American Veterinary Medical Association guidelines. Amniotic sacs were collected in either 10% formalin or fresh-frozen in liquid nitrogen and stored at −80°C until further analysis.

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