I am looking for a way to produce in vitro bovine embryos with good quality to make transcriptomic analysis of different genes related to elongation and gastrulation. Does anyone know an in vitro culture system?
Jack Rutledge of the Univ of Wisconsin did some day 10 to 14 embryo work -- maybe that will help you. http://www.ansci.wisc.edu/facstaff/Faculty/pages/rutledge/index.html
Hi Fernando, erythrocytes to embryo culture!? Do you have an explanation to do that?
Hi Jesica. The answer as far as I know is that no-one has successfully reproduced true elongation in vitro in a ruminant with the development and differentiation of the embryonic disc. There is some work on growing trophectoderm sacs, some of which includes constraining in tubes, so they "elongate", but my understanding is that there is no normal post-hatching in vitro. Our own observation is that from hatching, you see a lot of apotosis in the inner cell mass, until you simply end up with a trophoblastic vesicle. There are those which produce stem-cell - like cells with blastocysts planted onto feeder layers but again, this is not normal post-hatching development. Tom Spencer at Washington State Univ may also have some insights to resolving this problem. Good luck with this one!
Hello Jesica, I fully agree with Jeremy's comment, to my knowledge, there no example of physiological evolution of bovine embryos beyond hatching. Maybe you can see some gene expression changes as hatched blastocysts grow as round shaped vesicles in post hatching culture but not sure it has any physiological significance. Alternatively, it is possible to transfer blastocysts into transient recipient cows and flush them back at different times (up to Day 14-15) of elongation. Good luck.
Jessica, I agree with Pascal and Jeremy. You really would need to transfer into recipients on day 7 and then flush back at different times, or likely slaughter the recipients and flush the conceptuses back. We have done this type of work - look at
Forde N, Beltman ME, Duffy GB, Duffy P, Ó Gaora P, Roche JF, Lonergan P and Crowe MA (2011) Changes in the endometrial transcriptome during the bovine estrous cycle: effect of low circulating progesterone and consequences for conceptus elongation. Biology of Reproduction 84: 266-278.
and
Beltman ME, Lonergan P, Diskin MG, Roche JF and Crowe MA (2009). Effect of progesterone supplementation in the first week post conception on embryo survival in beef heifers. Theriogenology 71: 1173-1179.
These papers are available on my research gate page if you want the full text
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