Abstract
Spermatogonial stem cells (SSCs) are defined by unique properties like other stem cells. However, there are two major challenges: long-term cultivation of normal SSCs into stable cell lines and maintaining the SSCs as undifferentiated and capable of self-renewal. Here, we compared different culture methods for mouse SSCs isolated and cultured from testicular tissue. We found that human amniotic epithelial cells (hAECs) can behave as feeder cells, allowing mouse SSCs to maintain a high level of alkaline phosphatase (AP) activity when cultured long-term. Also, we observed that expression of Nanog, Oct-4 and other important stem cells markers were higher in mouse SSCs cultured on hAECs compared to those cultured on MEF or without any feeder cells. Furthermore, we demonstrated that the CpG islands of the Nanog and Oct-4 promoters were hypomethylated in cells cultured on hAECs. In addition, mouse SSCs cultured on hAECs exhibited higher levels of H3AC and H3K4Me3 in the Nanog and Oct-4 promoters than those cultured on MEF or without feeder cells. Taken together, these results suggest that the hAECinduced epigenetic modifications at the Nanog and Oct-4 locus could be a key mechanism for maintaining mouse SSCs in an undifferentiated state capable of self-renewal.
Article PDF
Similar content being viewed by others
References
Adams, G. B., Scadden, D. T. (2006) The hematopoietic stem cell in its place. Nat. Immunol. 7, 333–337.
Akle, C. A., Adinolfi, M., Welsh, K. I. (1981) Immunogenicity of human amniotic epithelial cells after transplantation into volunteers. Lancet 2, 1003–1005.
Bibikova, M., Chudin, E., Wu, B. (2006) Human embryonic stem cells have a unique epigenetic signature. Genome Res. 16, 1075–1083.
Boyer, L. A., Plath, K., Zeitlinger, J. (2006) Polycomb complexes repress developmental regulators in murine embryonic stem cells. Nature 441, 349–353.
Chambers, I., Colby, D., Robertson, M. (2003) Functional expression cloning of Nanog, a pluripotency sustaining factor in embryonic stem cells. Cell 113, 643–655.
Chen, B., Wang, Y. B., Zhang, Z. L. (2009) Xeno-free culture of human spermatogonial stem cells supported by human embryonic stem cell-derived fibroblast-like cells. Asian J. Androl. 11, 557–565.
Conrad, S., Renninger, M., Hennenlotter, J. (2008) Generation of pluripotent stem cells from adult human testis. Nature 456, 344–349.
Cowan, C. A., Atienza, J., Melton, D. A. (2005) Nuclear reprogramming of somatic cells after fusion with human embryonic stem cells. Science 309, 1369–1373.
de Rooij, D. G. (2001) Proliferation and differentiation of spermatogonial stem cells. Reproduction 121, 347–354.
Dym, M., Kokkinaki, M., He, Z. (2009) Spermatogonial stem cells: mouse and human comparisons. Birth Defects Res. Embryo Today 87, 27–34.
Dym, M., He, Z., Jiang, J. (2009) Spermatogonial stem cells: unlimited potential. Reprod. Fertil. Dev. 21, 15–21.
Freberg, C. T., Dahl, J. A., Timoskainen, S. (2007) Epigenetic reprogramming of OCT4 and NANOG regulatory regions by embryonal carcinoma cell extract. Mol. Biol. Cell 18, 1543–1553.
Giuili, G., Tomljenovic, A., Labrecque, N. (2002) Murine spermatogonial stem cells: targeted transgene expression and purification in an active state. EMBO Rep. 3, 753–759.
Guan, K., Nayernia, K., Maier, L. S. (2006) Pluripotency of spermatogonial stem cells from adult mouse testis. Nature 440, 1199–1203.
Hattori, N., Imao, Y., Nishino, K. (2007) Epigenetic regulation of Nanog gene in embryonic stem and trophoblast stem cells. Genes Cells 12, 387–396.
Hermann, B. P., Sukhwani, M., Simorangkir, D. R. (2009) Molecular dissection of the male germ cell lineage identifies putative spermatogonial stem cells in rhesus macaques. Hum. Reprod. 24, 1704–1716.
Hofmann, M. C., Braydich-Stolle, L., Dym, M. (2005) Isolation of male germ-line stem cells; influence of GDNF. Dev. Biol. 279, 114–124.
Hong, Y., Liu, T., Zhao, H. (2004) Establishment of a normal medakafish spermatogonial cell line capable of sperm production in vitro. Proc. Natl. Acad. Sci. USA 101, 8011–8016.
Izadyar, F., Pau, F., Marh, J. (2008) Generation of multipotent cell lines from a distinct population of male germ line stem cells. Reproduction 135, 771–784.
Kanatsu-Shinohara, M., Inoue, K., Lee, J. (2004) Generation of pluripotent stem cells from neonatal mouse testis. Cell 119, 1001–1012.
Kim, T. H., Barrera, L. O., Zheng, M. (2005) A high-resolution map of active promoters in the human genome. Nature 436, 876–880.
Kimura, H., Tada, M., Nakatsuji, N. (2004) Histone code modifications on pluripotential nuclei of reprogrammed somatic cells. Mol. Cell Biol. 24, 5710–5720.
Koizumi, N. J., Inatomi, T. J., Sotozono, C. J. (2000) Growth factor mRNA and protein in preserved human amniotic membrane. Curr. Eye Res. 20, 173–177.
Lachner, M., Jenuwein, T. (2002) The many faces of histone lysine methylation. Curr. Opin. Cell Biol. 14, 286–298.
Lai, D., Cheng, W., Liu, T. (2009) Use of Human Amnion Epithelial Cells as a Feeder Layer to Support Undifferentiated Growth of Mouse Embryonic Stem Cells. Cloning Stem Cells 11, 331–340.
Lee, T. I., Jenner, R. G., Boyer, L. A. (2006) Control of developmental regulators by Polycomb in human embryonic stem cells. Cell 125, 301–313.
Loh, Y. H., Wu, Q., Chew, J. L. (2006) The Oct4 and Nanog transcription network regulates pluripotency in mouse embryonic stem cells. Nat. Genet. 38, 431–440.
Lucas, B., Fields, C., Hofmann, M. C. (2009) Signaling pathways in spermatogonial stem cells and their disruption by toxicants. Birth Defects Res. Embryo Today 87, 35–42.
Maki, C. B., Pacchiarotti, J., Ramos, T. (2009) Phenotypic and molecular characterization of spermatogonial stem cells in adult primate testes. Hum. Reprod. 24, 1480–1491.
Meng, X., Lindahl, M., Hyvonen, M. E. (2000) Regulation of cell fate decision of undifferentiated spermatogonia by GDNF. Science 287, 1489–1493.
Mitsui, K., Tokuzawa, Y., Itoh, H. (2003) The homeoprotein Nanog is required for maintenance of pluripotency in mouse epiblast and ES cells. Cell 113, 631–642.
Miyabayashi, T., Teo, J. L., Yamamoto, M. (2007) Wnt/beta-catenin/CBP signaling maintains longterm murine embryonic stem cell pluripotency. Proc. Natl. Acad. Sci. USA 104, 5668–5673.
Niwa, H., Miyazaki, J., Smith, A. G. (2000) Quantitative expression of Oct-3/4 defines differentiation, dedifferentiation or self-renewal of ES cells. Nat. Genet. 24, 372–376.
Seandel, M., James, D., Shmelkov, S. V. (2007) Generation of functional multipotent adult stem cells from GPR125+ germline progenitors. Nature 449, 346–350.
Shinohara, T., Brinster, R. L. (2000) Enrichment and transplantation of spermatogonial stem cells. Int. J. Androl. 23 Suppl 2, 89–91.
Shinohara, T., Avarbock, M. R., Brinster, R. L. (1999) beta1- and alpha6-integrin are surface markers on mouse spermatogonial stem cells. Proc. Natl. Acad. Sci. USA 96, 5504–5509.
Simonsson, S., Gurdon, J. (2004) DNA demethylation is necessary for the epigenetic reprogramming of somatic cell nuclei. Nat. Cell. Biol. 6, 984–990.
Tada, M., Tada, T., Lefebvre, L. (1997) Embryonic germ cells induce epigenetic reprogramming of somatic nucleus in hybrid cells. EMBO J. 16, 6510–6520.
Takahashi, K., Yamanaka, S. (2006) Induction of pluripotent stem cells from mouse embryonic and adult fibroblast cultures by defined factors. Cell. 126, 663–676.
Wu, Z., Falciatori, I., Molyneux, L. A. (2009) Spermatogonial culture medium: an effective and efficient nutrient mixture for culturing rat spermatogonial stem cells. Biol. Reprod. 81, 77–86.
Xie, T., Spradling, A. C. (2000) A niche maintaining germ line stem cells in the Drosophila ovary. Science 290, 328–330.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made.
About this article
Cite this article
Liu, T., Huang, Y., Huang, Q. et al. Use of Human Amniotic Epithelial Cells as a Feeder Layer to Support Undifferentiated Growth of Mouse Spermatogonial Stem Cells Via Epigenetic Regulation of the Nanog and Oct-4 Promoters. BIOLOGIA FUTURA 63, 167–179 (2012). https://doi.org/10.1556/ABiol.63.2012.2.1
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1556/ABiol.63.2012.2.1