Ratio of inner cell mass and trophoblastic cells in blastocysts derived from porcine 4- and 8-cell embryos and isolated blastomeres cultured in vitro in the presence or absence of protein and human leukemia inhibitory factor
- PMID: 9282990
- DOI: 10.1095/biolreprod57.3.552
Ratio of inner cell mass and trophoblastic cells in blastocysts derived from porcine 4- and 8-cell embryos and isolated blastomeres cultured in vitro in the presence or absence of protein and human leukemia inhibitory factor
Abstract
In this study we investigated effects of developmental stage at onset of individual in vitro culture on the progress of development in intact embryos and individual blastomeres derived from 4-cell and 8-cell porcine embryos (referred to as 1/4 and 1/8, respectively), the necessity of serum or BSA supplementation in culture media for embryos and blastomeres (in contrast to development in a defined medium), and the role of two concentrations of human leukemia inhibitory factor (hLIF) on development of blastomeres and embryos. More (p < 0.05) 1/4 blastomeres developed to the blastocyst stage than did 1/8 blastomeres. In the serum-supplemented medium, the percentage of inner cell mass (ICM)/total cells in 8-cell- and 1/8-derived blastocysts was higher (p < 0.05) than that in 4-cell- and 1/4-derived embryos. Development to blastocysts was similar in BSA-supplemented and defined medium as compared to that in serum-enriched medium in intact 4-cell and 8-cell embryos, and 1/4 blastomeres. More (p < 0.05) 1/8 blastomeres developed to the blastocyst stage in serum-supplemented medium than in defined medium. The high hLIF concentration (1000 IU/ml) decreased (p < 0.05) blastocyst development in 1/4 blastomeres in defined medium, but fewer blastocysts (p > 0.05) lacked an ICM (blastocyst-like vesicles) than in defined medium without hLIF. It is concluded that 1) porcine intact embryos and isolated blastomeres can be cultured individually in defined medium up to the blastocyst stage from the 4-cell stage onwards; 2) more 1/4 isolated blastomeres develop to blastocysts with more total cells but with a lower ratio of ICM to total cells than blastocysts derived from 1/8 blastomeres; 3) the effects of hLIF are dependent on proteins present in the culture medium and on the embryonic stage; and 4) in defined medium, high concentrations of hLIF are inhibitory to blastocyst formation, but fewer blastocyst-like vesicles are formed. The defined culture system employed in this study allows examination of the effects of growth factors or cytokines in porcine early embryonic development.
Similar articles
-
Cellular characterization of blastocysts derived from rabbit 4-, 8- and 16-cell embryos and isolated blastomeres cultured in vitro.Hum Reprod. 2000 Apr;15(4):881-9. doi: 10.1093/humrep/15.4.881. Hum Reprod. 2000. PMID: 10739837
-
Culture of bovine embryos in buffalo rat liver cell-conditioned media or with leukemia inhibitory factor.J Anim Sci. 1997 May;75(5):1332-6. doi: 10.2527/1997.7551332x. J Anim Sci. 1997. PMID: 9159281
-
Effects of gas atmosphere, platelet-derived growth factor and leukemia inhibitory factor on cell numbers of rabbit embryos cultured in a protein-free medium.Reprod Nutr Dev. 1997;37(1):97-104. doi: 10.1051/rnd:19970110. Reprod Nutr Dev. 1997. PMID: 9115600
-
Progress in reproductive biotechnology in swine.Theriogenology. 2001 Nov 1;56(8):1291-304. doi: 10.1016/s0093-691x(01)00630-6. Theriogenology. 2001. PMID: 11758883 Review.
-
The effect of leukaemia inhibitory factor (LIF) on embryogenesis.Reprod Fertil Dev. 1992;4(4):449-58. doi: 10.1071/rd9920449. Reprod Fertil Dev. 1992. PMID: 1461995 Review.
Cited by
-
Effect of different concentrations of recombinant leukemia inhibitory factor on different development stage of mouse embryo in vitro.J Assist Reprod Genet. 2000 Jul;17(6):352-5. doi: 10.1023/a:1009413329977. J Assist Reprod Genet. 2000. PMID: 11042834 Free PMC article.
-
Modulation of pluripotency in the porcine embryo and iPS cells.PLoS One. 2012;7(11):e49079. doi: 10.1371/journal.pone.0049079. Epub 2012 Nov 8. PLoS One. 2012. PMID: 23145076 Free PMC article.
-
Improved Murine Blastocyst Quality and Development in a Single Culture Medium Compared to Sequential Culture Media.Reprod Sci. 2016 Mar;23(3):310-7. doi: 10.1177/1933719115618281. Epub 2015 Dec 13. Reprod Sci. 2016. PMID: 26668049 Free PMC article.
-
Developmental potential of isolated blastomeres from early mouse embryos in the presence and absence of LIF and GM-CSF.J Assist Reprod Genet. 2008 Jan;25(1):7-12. doi: 10.1007/s10815-007-9191-0. Epub 2008 Jan 18. J Assist Reprod Genet. 2008. PMID: 18202911 Free PMC article.
-
Identification and isolation of embryonic stem cells in reproductive endocrinology: theoretical protocols for conservation of human embryos derived from in vitro fertilization.Theor Biol Med Model. 2005 Jul 18;2:25. doi: 10.1186/1742-4682-2-25. Theor Biol Med Model. 2005. PMID: 16026616 Free PMC article.
Publication types
MeSH terms
Substances
LinkOut - more resources
Full Text Sources