A Laboratory Guide to the Mammalian Embryo by David K. Gardner, Michelle Lane, Andrew J. Watson

By David K. Gardner, Michelle Lane, Andrew J. Watson

This publication pulls jointly the total variety of mobilephone tradition, biochemical, microscopic, and genetic suggestions to check the early mammalian embryo. previously, there hasn't ever been one of these entire compendium, even though there were extra centred books of protocol, corresponding to Manipulating the Mouse Embryo, from chilly Spring Harbor. This booklet is meant to attract all constituencies, from uncomplicated experimental technological know-how to medical and animal technology functions.

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The development of recombinant human serum albumin has therefore helped eliminate the problems inherent with using blood-derived products and should eliminate variability. Albumin is the most abundant protein in the female reproductive tract (16); however, there are other macromolecules, such as mucins, that are present at higher levels than albumin (29). The role of mucins during the preimplantation period has yet to be elucidated, but their role during implantation has been better characterized (30).

It has been reported that using mice between 4 and 5 weeks old results in a low yield of quality oocytes (19). 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.

Locate cumulus-enclosed oocytes from the follicular aspirate. A grid is scratched onto the outer bottom of a 10-mm petri dish using a scalpel blade or needle to assist in the searching. Using a sterile Pasteur pipette, remove the bottom 1 ml of the follicular aspirate containing the sediment from the tube and transfer it to the searching plate. To this sediment add 1–2 ml of HEPES-TCM-199 medium. Add a further 2–3 ml of HEPES-TCM-199 medium to a 35-mm petri dish for collection of the cumulus-enclosed oocyte complexes.

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