Date of Award

1990

Document Type

Dissertation

Degree Name

Doctor of Philosophy (PhD)

Department

School of Animal Science

First Advisor

Robert A. Godke

Abstract

A novel embryo culture system has been developed using 96-hour chick embryos. One to four mammalian embryos can be injected into the chick embryo amnion (CEA) and allowed to develop for 72 to 96 hours. Pronuclear-stage mouse embryos from two different strains were cultured in the CEA or in Whitten's medium. There were more expanded blastocysts from one strain of embryos when cultured in the CEA. More hatched blastocysts resulted from embryos of both strains when cultured in the CEA. Two to eight-cell goat embryos cultured in the CEA for 72 hours or on cell monolayers reached to blastocyst stage at higher rates than when cultured with trophoblastic vesicles or in medium alone. When culture in the CEA was extended to 96 hours, more blastocyst were obtained than when embryos were co-cultured on monolayers for 96 hours or in medium alone. More expanded blastocysts were observed following the culture of precompaction stage bovine morulae in the CEA than when embryos were cultured on monolayers or cultured in medium alone. Culture of bovine morulae on monolayers or in the CEA prior to freezing improved post-thaw viability when compared with culture in medium alone. When in vitro fertilization (IVF)-derived bovine embryos were cultured sequentially in two or three CEA, development was not improved over culture with cumulus cells and unacceptable loss of embryos occurred. The culture of IVF-derived embryos in the chick embryos during the first 48 hours of development resulted in less four to six-cell embryos than culture with cumulus cells, however, culture of later-stage IVF-derived embryos in the CEA appears to be as effective as cumulus cell co-culture. Extracted chick amniotic fluids (CAF) were used to supplement the culture medium for mouse and cow embryos. Two-cell mouse embryos developed at similar rates when cultured in CAF or fetal bovine serum (FBS) supplemented medium, however, embryos placed in the CEA cleaved at higher rates. The use of CAF as a supplement in in vitro maturation and culture medium for bovine IVF procedures appears to be as effective as supplementation with FBS.

Pages

156

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