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Title: | Pregnancy rate and survival in culture of in vitro fertilized bovine embryos frozen in various cryoprotectants and thawed using a one-step system |
Authors: | Suzuki, Tatsuyuki M. Takagi M. Yamamoto Saha. S. Boediono, Arief Ooe, Masato |
Issue Date: | 1993 |
Publisher: | IPB (Bogor Agricultural University) |
Abstract: | Bovine oocytes surrounded with compact cumulus cells were cultured for 20-22 h (38.5°C, 5 % C02) in modified TCM -199 supplemented with 5 % superovulated cow serum (SCS) and inseminated by in vitro capacitated sperm. Day 7-8 embryos were equilibrated 10 min in 1.3 M methyl cellosolve (MC), 1.1 M diethylene glycol (DEG), 1.8 M ethylene glycol (EG), 1.6 M propylene glycol (PG) and 1.1 M l, 3-butylene glycol(BG) solutions, loaded into 0.25ml straws, placed into an alcohol bath freezer at 0°C, cooled from 0°C to -6°C at -l°C/min, seeded, held for 10 min, cooled at -0.3°C/min or -0.5°C/min to -30°C. Straws were then plunged and stored in liquid nitrogen. After thaw~ng in 30°C water, embryos were rehydrated directly in TCM-199 and then cultured for 48 hours in TCM-199 plus 5% SCS. Embryos were considered viable if they progressed to later developmental stages with good morphology. Some of the embryos frozen in each cryoprotectant were thawed and transferred non-surgically without removing the cryoprotectant. Hatched embryos survived freezing and one step dilution as follows: EG (50.0%), MC (53.6%), DEG (56.9%), PG (58.0%) and BG (11.5%). The survival rate of embryos cooled at 0.3°C/min versus 0.5°C/min was not significantly different (P>0.05). However, blastocysts hatched most often (P< 0.01) in vitro when cooled at a rate of 0.3°C/min [64.6 %(31/48)] compared with 0.5°C/min [22.6% (12 /53)]. Embryo survival in TCM-199 culture relative to cryoprotectant is shown in the table below. Pregnancy rates resulting from embryos frozen in different cryoprotectants were as follows: MC(48%, 10/21), DEG(30%, 3/10), EG(87%, 20 23), and PG(40%, 4/10). |
URI: | http://repository.ipb.ac.id/handle/123456789/53232 |
Appears in Collections: | Anatomy, Phisiology and Pharmacology |
Files in This Item:
File | Description | Size | Format | |
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Therio1993-39-324-PregFrezzAbs.pdf | Publication | 54 kB | Adobe PDF | View/Open |
ABO_Pregnancy rate and survival_Abstract.pdf | Abstract | 44.05 kB | Adobe PDF | View/Open |
ABO_Pregnancy rate and survival_Abstract.doc | Abstract | 28.5 kB | Microsoft Word | View/Open |
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