By David K. Gardner, Michelle Lane, Andrew J. Watson
This booklet pulls jointly the whole variety of mobile tradition, biochemical, microscopic, and genetic innovations to check the early mammalian embryo. formerly, there hasn't ever been the sort of finished compendium, even though there were extra concentrated books of protocol, equivalent to Manipulating the Mouse Embryo, from chilly Spring Harbor. This booklet is meant to attract all constituencies, from simple experimental technology to medical and animal technological know-how functions.
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Additional resources for A Laboratory Guide to the Mammalian Embryo
Oocytes surrounded by granulosa cells are aspirated from ovaries 44–46 h after PMSG injection. Ovaries are removed from the female in warmed 28 A LABORATORY GUIDE TO THE MAMMALIAN EMBRYO handling media. 6). Once mouse oocytes are removed from the follicles, the zona hardens and becomes impenetrable by sperm. Therefore it is necessary to either include serum in the media for collection or add fetuin (1 mg/ml) to media without serum. 7). The success of IVF in the mouse is highly dependent on the strain of male mouse.
Mol. Reprod. , 54, 410. 4. Ludwig, T. , Squirrell, J. , Palmenberg, A. , and Bavister, B. D. (2001). Biol. , 65, 1648. 5. Schini, S. , and Bavister, B. D. (1988). Biol. , 39, 1183. 6. Seshagiri, P. , and Bavister, B. D. (1991). Mol. Reprod. , 30, 105. 7. Quinn, P. (1995). J. Assist. Reprod. , 12, 97. 8. , Steinberg, J. , and Weathersbee, P. S. (1995). Fertil. , 63, 922. 9. Gardner, D. , and Lane, M. (1997). Hum. Reprod. Update, 3, 367. 10. Gardner, D. , and Lane, M. (1998). Hum. , 13 Suppl. 3, 148.
3, as this is the physiological intracellular pH for mammalian embryos. This is particularly important for the manipulation of oocytes, as they appear to lack any active pH transport systems (12–14). Handling media are usually supplemented with serum albumin (BSA, HSA) or a synthetic macromolecule such as PVA. The use of serum should be avoided due to its detrimental actions on embryo physiology (10, 15–18). 2 Quality Control The quality of the media used for handling gametes and embryos depends on the quality of the components used to prepare the media.
A Laboratory Guide to the Mammalian Embryo by David K. Gardner, Michelle Lane, Andrew J. Watson