Improved development of rabbit one-cell embryos to the hatching blastocyst stage by culture in a defined, protein-free culture medium
| Autor: | E. W. Carney, R H Foote |
|---|---|
| Rok vydání: | 1991 |
| Předmět: |
Embryology
Serum albumin Biology Andrology Embryonic and Fetal Development Selenium Endocrinology medicine Animals Insulin Blastocyst Bovine serum albumin Fetal Viability Zona pellucida Epidermal Growth Factor Hatching Embryogenesis Transferrin Obstetrics and Gynecology Serum Albumin Bovine Embryo Embryo culture Cell Biology Culture Media medicine.anatomical_structure Reproductive Medicine Polyvinyl Alcohol embryonic structures Immunology biology.protein Rabbits |
| Zdroj: | Reproduction. 91:113-123 |
| ISSN: | 1741-7899 1470-1626 |
| DOI: | 10.1530/jrf.0.0910113 |
| Popis: | In Exp. 1, Medium 199 and Medium RD (RPMI-1640 and Dulbecco's MEM, 1:1 v/v) were compared in a 2 x 2 factorial design by supplementing each with 15 mg bovine serum albumin (BSA)/ml of 1 mg polyvinyl alcohol (PVA)/ml. All media contained 5 micrograms insulin/ml, 5 micrograms transferrin/ml, 5 ng selenium/ml (ITS), and 10 ng epidermal growth factor (EGF)/ml. One-cell embryos were cultured at 39 degrees C with 5% CO2 in air for 65 h and then stained with Hoechst 33342 to determine blastomere number. Embryos in Medium 199 developed poorly (P less than 0.001) when PVA was used instead of BSA (30 vs 76 cells/embryo), but developed rapidly in Medium RD with PVA or BSA (118 and 121 cells). Similar results were obtained in Exp. 2 in BSA- and PVA-free medium. In Exp. 3, the development of 1-cell embryos after 65 h in unsupplemented (protein-free) Medium RD (68% blastocysts, 117 cells) did not differ (P greater than 0.37) from that obtained using Medium RD with insulin, ITS or EGF alone. Culture in protein-free Medium RD for 96 h resulted in 82% of the 1-cell embryos forming blastocysts and 40% hatching through the zona pellucida. In a preliminary test of viability, 1-cell embryos cultured in this medium for 48 or 65 h and transferred to synchronous recipients resulted in 5/18 (28%) and 3/24 (12%) Day-15 viable fetuses. Cell counts of approximately 120 per blastocyst after culturing 1-cell embryos for 65 h in Medium RD indicated that cell division was more rapid than that obtained with all other media tested previously in this laboratory. This is the first report of rabbit embryo development from the 1-cell to the hatching blastocyst stage in a defined protein-free culture medium. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|