Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2012 Oct;29(10):1021-8.
doi: 10.1007/s10815-012-9824-9. Epub 2012 Jun 29.

Co-localization of NANOG and OCT4 in human pre-implantation embryos and in human embryonic stem cells

Fredwell Hambiliki  1 , Susanne Ström, Pu Zhang, Anneli Stavreus-Evers
Affiliations

Co-localization of NANOG and OCT4 in human pre-implantation embryos and in human embryonic stem cells

Fredwell Hambiliki et al. J Assist Reprod Genet. 2012 Oct.

Abstract

Purpose: NANOG and OCT4 are required for the maintenance of pluripotency in embryonic stem cells (ESCs). These proteins are also expressed in the inner cell mass (ICM) of the mouse pre-implantation embryo.

Methods: Immunohistochemistry was used to show the presence of NANOG and OCT4 protein, and in situ hybridization was used to localize NANOG mRNA in human embryos from two-cell to blastocyst stage, and in human ESCs (hESCs).

Results: Nanog and Oct4 were co-localized in human embryos from morula and blastocyst stages. NANOG mRNA was detected in a group of cells in the morula, in cells of the ICM of blastocysts, and evenly in hESCs. All non-differentiated hESCs expressed NANOG and OCT4 protein. Pluripotent cells expressing NANOG and Oct4 were eccentrically localized, probably in polarized cells in a human compacted morula, which appears to be different from expression in murine embryos.

Conclusion: In this study, we demonstrate that whole mount in situ hybridization is amenable to localization of mRNAs in human development, as in other species.

PubMed Disclaimer

Figures

Fig. 1
Fig. 1
Representative pictures show double staining of OCT4 and NANOG in the human morula and blastocyst. a Immunostaining of OCT4 in the morula. b Immunostaining of NANOG in the same morula. c DAPI staining in the morula. d Staining of OCT4 in the blastocyst. Immunostaining is seen both in the inner cell mass and the trophoblast; arrows. e Staining of NANOG in the same blastocyst. Staining is seen only in the inner cell mass; arrow. f DAPI staining in the same blastocyst. g Exclusion of OCT4 antibody. h Exclusion of NANOG antibody. i DAPI staining of the embryo without primary antibodies present
Fig. 2
Fig. 2
In situ hybridization of NANOG in the human pre-implantation embryo. a 2-cell embryo, day 1. b 4-cell embryo, day 2. c 8-cell embryo, day 3. d morula, day 4. e late day 4 embryo. f blastocyst, day 5. g mouse ESCs hybridized with human NANOG primer
Fig. 3
Fig. 3
Immunostaining of OCT4 and NANOG in human embryonic stem cells, and in situ hybridization of NANOG in embryonic stem cells. a Light microscopy of human embryonic stem cells. b NANOG immunostaining in embryonic stem cells. The feeder cells do not show staining for NANOG (red arrow). c Nuclear staining of embryonic stem cells and fibroblast cells. DAPI staining is seen in feeder cells (blue arrow). d & e In situ hybridization showing NANOG mRNA in embryonic stem cells. NANOG f Mouse embryonic stem cells hybridized with human NANOG primer. g OCT4 immunostaining of stem cell line HS426 (green). Blue colour shows nuclear DAPI staining
See this image and copyright information in PMC

References

    1. Bjuresten K, Hovatta O. Donation of embryos for stem cell research—how many couples consent? Hum Reprod. 2003;18:1353–1355. doi: 10.1093/humrep/deg265. - DOI - PubMed
    1. Buehr M, Smith A. Genesis of embryonic stem cells. Philos Trans R Soc Lond B Biol Sci. 2003;358:1397–4402. doi: 10.1098/rstb.2003.1327. - DOI - PMC - PubMed
    1. Chambers I, Colby D, Robertson M, Nichols J, Lee S, Tweedie S, Smith A. Functional expression cloning of NANOG, a pluripotency sustaining factor in Embronic Stem cells. Cell. 2003;113:643–655. doi: 10.1016/S0092-8674(03)00392-1. - DOI - PubMed
    1. Chambers I, Smith A. Self renewal of teratocarcinoma and embryonic stem cells. Oncogene. 2004;23:7150–7160. doi: 10.1038/sj.onc.1207930. - DOI - PubMed
    1. Puy L, Lopes SM, Haagsman HP, Roelen BA. Analysis of co-expression of OCT4, NANOG and SOX2 in pluripotent cells of the porcine embryo, in vivo and in vitro. Therienology. 2011;75:513–526. doi: 10.1016/j.theriogenology.2010.09.019. - DOI - PubMed

Publication types

MeSH terms