Skip to main content
SpringerLink
Log in

Expression of TGFβ family factors and FGF2 in mouse and human embryonic stem cells maintained in different culture systems

  • Developmental Biology of Mammals
  • Published:
Russian Journal of Developmental Biology Aims and scope Submit manuscript

Abstract

Mouse and human embryonic stem cells are in different states of pluripotency (naive/ground and primed states). Mechanisms of signaling regulation in cells with ground and primed states of pluripotency are considerably different. In order to understand the contribution of endogenous and exogenous factors in the maintenance of a metastable state of the cells in different phases of pluripotency, we examined the expression of TGFβ family factors (ActivinA, Nodal, Lefty1, TGFβ1, GDF3, BMP4) and FGF2 initiating the appropriate signaling pathways in mouse and human embryonic stem cells (mESCs, hESCs) and supporting feeder cells. Quantitative real-time PCR analysis of gene expression showed that the expression patterns of endogenous factors studied were considerably different in mESCs and hESCs. The most significant differences were found in the levels of endogenous expression of TGFβ1, BMP4 and ActivinA. The sources of exogenous factors ActivnA, TGFβ1, and FGF2 for hESCs are feeder cells (mouse and human embryonic fibroblasts) expressing high levels of these factors, as well as low levels of BMP4. Thus, our data demonstrated that the in vitro maintenance of metastable state of undifferentiated pluripotent cells is achieved in mESCs and hESCs using different schemes of the regulations of ActivinA/Nodal/Lefty/Smad2/3 and BMP/Smad1/5/8 endogenous branches of TGFβ signaling. The requirement for exogenous stimulation or inhibition of these signaling pathways is due to different patterns of endogenous expression of TGFβ family factors and FGF2 in the mESCs and hESCs. For the hESCs, enhanced activity of ActivinA/Nodal/Lefty/Smad2/3 signaling by exogenous factor stimulation is necessary to mitigate the effects of BMP/Smad1/5/8 signaling pathways that promote cell differentiation into the extraembryonic structures. Significant differences in endogenous FGF2 expression in the cells in the ground and primed states of pluripotency demonstrate diverse involvement of this factor in the regulation of the pluripotent cell self-renewal.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Beattie, G.M., Lopez, A.D., Bucay, N., et al., Activin A Maintains Pluripotency of Human Embryonic Stem Cells in the Absence of Feeder Layers, Stem Cells, 2005, vol. 23, pp. 489–495.

    Article  PubMed  CAS  Google Scholar 

  • Becker, K.A., Ghule, P.N., Therrien, J.A., et al., Self-Renewal of Human Embryonic Stem Cells Is Supported by a Shortened G1 Cell Cycle Phase, J. Cell Physiol., 2006, vol. 209, pp. 883–893.

    Article  PubMed  CAS  Google Scholar 

  • Brons, I.G.M., Smithers, L.E., Trotter, M.W.B., et al., Derivation of Pluripotent Epiblast Stem Cells from Mammalian Embryos, Nature, 2007, vol. 448, pp. 191–195.

    Article  PubMed  CAS  Google Scholar 

  • Burdon, T., Smith, A., and Savatier, P., Signaling, Cell Cycle and Pluripotency in Embryonic Stem Cells, Trends Cell Biol., 2002, vol. 12, pp. 432–438.

    Article  PubMed  CAS  Google Scholar 

  • Dahéron, L., Opitz, S.L., Zaehres, H., et al., LIF/STAT3 Signaling Fails to Maintain Self-Renewal of Human Embryonic Stem Cells, Stem Cells, 2004, vol. 22, pp. 770–778.

    Article  PubMed  Google Scholar 

  • Dreesen, O. and Brivanlou, A.H., Signaling Pathways in Cancer and Embryonic Stem Cells, Stem Cell Rev., 2007, vol. 3, pp. 7–17.

    Article  PubMed  CAS  Google Scholar 

  • Dunn, N.R., Vincent, S.D., Oxburgh, L., et al., Combinatorial Activities of Smad2 and Smad3 Regulate Mesoderm Formation and Patterning in the Mouse Embryo, Development, 2004, vol. 131, pp. 1717–1728.

    Article  PubMed  CAS  Google Scholar 

  • Eiselleova, L., Peterkova, I., Neradil, J., et al., Comparative Study of Mouse and Human Feeder Cells for Human Embryonic Stem Cells, Int. J. Dev. Biol., 2008, vol. 52, pp. 353–363.

    Article  PubMed  CAS  Google Scholar 

  • Eiselleova, L., Matulka, K., Kriz, V., et al., A Complex Role for FGF-2 in Self-Renewal, Survival, and Adhesion of Human Embryonic Stem Cells, Stem Cells, 2009, vol. 27, pp. 1847–1857.

    Article  PubMed  CAS  Google Scholar 

  • Evseenko, D., Schenke-Layland, K., Dravid, G., et al., Identification of the Critical Extracellular Matrix Proteins that Promote Human Embryonic Stem Cell Assembly, Stem Cells Dev., 2009, vol. 18, pp. 919–928.

    Article  PubMed  CAS  Google Scholar 

  • Gordeeva, O.F., Lifantseva, N.V., and Khaidukov, S.V., Expression Patterns of Germ Line Specific Genes in Mouse and Human Pluripotent Stem Cells Are Associated with Regulation of Ground and Primed State of Pluripotency, Russ. J. Dev. Biol., 2011, vol. 42, no. 6, pp. 355–375.

    Article  CAS  Google Scholar 

  • Greber, B., Lehrach, H., and Adjaye, J., Fibroblast Growth Factor 2 Modulates Transforming Growth Factor β Signaling in Mouse Embryonic Fibroblasts and Human ESCs (hESCs) to Support hESC Self-Renewal, Stem Cells, 2007, vol. 25, pp. 455–464.

    Article  PubMed  CAS  Google Scholar 

  • Guo, G., Yang, J., Nichols, J., et al., Klf4 Reverts Developmentally Programmed Restriction of Ground State Pluripotency, Development, 2009, vol. 136, pp. 1063–1069.

    Article  PubMed  CAS  Google Scholar 

  • Han, D.W., Tapia, N., Joo, J.Y., et al., Epiblast Stem Cell Subpopulations Represent Mouse Embryos of Distinct Pregastrulation Stages, Cell, 2010, vol. 143, pp. 617–627.

    Article  PubMed  CAS  Google Scholar 

  • Hanna, J., Cheng, A.W., Saha, K., et al., Human Embryonic Stem Cells with Biological and Epigenetic Characteristics Similar to Those of Mouse ESCs, Proc. Natl. Acad. Sci. USA, 2010, vol. 107, pp. 9222–9227.

    Article  PubMed  CAS  Google Scholar 

  • Hayashi, K. and Surani, M.A., Resetting the Epigenome Beyond Pluripotency in the Germline, Cell Stem Cell, 2009, vol. 4, pp. 493–498.

    Article  PubMed  CAS  Google Scholar 

  • James, D., Levine, A.J., Besser, D., and Hemmati-Brivanlou, A., TGFbeta/Activin/Nodal Signaling Is Necessary for the Maintenance of Pluripotency in Human Embryonic Stem Cells, Development, 2005, vol. 132, pp. 1273–1282.

    Article  PubMed  CAS  Google Scholar 

  • Kol’tsova, A.M., Gordeeva, O.F., Krylova, T.A., et al., Comparative Characteristics of New Human Embryonic Stem Cell Lines SC5, SC6, SC7, and SC3a, Russ. J. Dev. Biol., 2011, vol. 42, no. 4, pp. 212–225.

    Article  Google Scholar 

  • Kol’tsova, A.M., Voronkina, I.V., Gordeeva, O.F., et al., Developing of a New Feeder-Free System and Characterization of Human Embryonic Stem Cell Sublines Derived in This System under Autogenic and Allogenic Culturing, Tsitologiia, 2012, vol. 54, no. 8, pp. 637–651.

    PubMed  Google Scholar 

  • Lanner, F. and Rossant, J., The Role of FGF/Erk Signaling in Pluripotent Cells, Development, 2010, vol. 137, pp. 3351–3360.

    Article  PubMed  CAS  Google Scholar 

  • McLean, A.B., D’Amour, K.A., Jones, K.L., et al., Activin A Efficiently Specifies Definitive Endoderm from Human Embryonic Stem Cells Only when Phosphatidylinositol 3-Kinase Signaling Is Suppressed, Stem Cells, 2007, vol. 25, pp. 29–38.

    Article  PubMed  CAS  Google Scholar 

  • Montes, R., Ligero, G., Sanchez, L., et al., Feeder-Free Maintenance of hESCs in Mesenchymal Stem Cell-Conditioned Media: Distinct Requirements for TGF-beta and IGF-II, Cell Res., 2009, vol. 19, pp. 698–709.

    Article  PubMed  CAS  Google Scholar 

  • Mummery, C.L., Transforming Growth Factor Beta and Mouse Development, Microsc. Res. Techn., 2001, vol. 52, pp. 374–386.

    Article  CAS  Google Scholar 

  • Na, J., Furue, M.K., and Andrews, P.W., Inhibition of ERK1/2 Prevents Neural and Mesendodermal Differentiation and Promotes Human Embryonic Stem Cell Self-Renewal, Stem Cell Res., 2010, vol. 5, pp. 157–169.

    Article  PubMed  CAS  Google Scholar 

  • Nichols, J. and Smith, A., Naive and Primed Pluripotent States, Cell Stem Cell, 2009, vol. 4, pp. 487–492.

    Article  PubMed  CAS  Google Scholar 

  • Puce-at, M., TGFβ in the Differentiation of Embryonic Stem Cells, Cardiovasc. Res., 2007, vol. 74, pp. 256–261.

    Article  CAS  Google Scholar 

  • Savatier, P., Lapillonne, H., van Grunsven, L.A., et al., Withdrawal of Differentiation Inhibitory Activity/Leukemia Inhibitory Factor Up-Regulates D Type Cyclins and Cyclin-Dependent Kinase Inhibitors in Mouse Embryonic Stem Cells, Oncogene, 1996, vol. 12, pp. 309–322.

    PubMed  CAS  Google Scholar 

  • Silva, J., Barrandon, O., Nichols, J., et al., Promotion of Reprogramming to Ground State Pluripotency by Signal Inhibition, PLoS Biol., 2008, vol. 6, p. e253.

    Article  PubMed  Google Scholar 

  • Smith, A.G., Heath, J.K., Donaldson, D.D., et al., Inhibition of Pluripotential Embryonic Stem Cell Differentiation by Purified Polypeptides, Nature, 1988, vol. 336, pp. 688–690.

    Article  PubMed  CAS  Google Scholar 

  • Stojkovic, P., Lako, M., Stewart, R., et al., Stem Cells, 2005, vol. 23, pp. 306–314.

    Article  PubMed  CAS  Google Scholar 

  • Tesar, P.J., Chenoweth, J.G., Brook, F.A., et al., New Cell Lines from Mouse Epiblast Share Defining Features with Human Embryonic Stem Cells, Nature, 2007, vol. 448, pp. 196–199.

    Article  PubMed  CAS  Google Scholar 

  • Thomson, J.A., Itskovitz-Eldor, J., Shapiro, S.S., et al., Embryonic Stem Cell Lines Derived from Human Blastocysts, Science, 1998, vol. 282, pp. 1145–1147.

    Article  PubMed  CAS  Google Scholar 

  • Valdimarsdottir, G. and Mummery, C., Functions of the TGFb Superfamily in Human Embryonic Stem Cells, APMIS, 2005, vol. 113, pp. 773–789.

    Article  PubMed  CAS  Google Scholar 

  • Vallier, L., Alexander, M., and Pedersen, R.A., Activin/Nodal and FGF Pathways Cooperate to Maintain Pluripotency of Human Embryonic Stem Cells, J. Cell Sci., 2005, vol. 118, pp. 4495–4509.

    Article  PubMed  CAS  Google Scholar 

  • Vallier, L., Touboul, T., Chng, Z., et al., Early Cell Fate Decisions of Human Embryonic Stem Cells and Mouse Epiblast Stem Cells Are Controlled by the Same Signalling Pathways, PLoS ONE, 2009, vol. 4, p. e6082.

  • Xiao, L., Yuan, X., and Sharkis, S.J., Activin A Maintains Self-Renewal and Regulates Fibroblast Growth Factor, Wnt, and Bone Morphogenic Protein Pathways in Human Embryonic Stem Cells, Stem Cells, 2006, vol. 24, pp. 1476–1486.

    Article  PubMed  CAS  Google Scholar 

  • Xu, C., Inokuma, M.S., Denham, J., et al., Feeder-Free Growth of Undifferentiated Human Embryonic Stem Cells, Nature Biotechnol., 2001, vol. 19, pp. 971–974.

    Article  CAS  Google Scholar 

  • Xu, R.H., Chen, X., Li, D., et al., BMP4 Initiates Human Embryonic Stem Cell Differentiation to Trophoblast, Nat. Biotechnol., 2002, vol. 20, pp. 1261–1264.

    Article  PubMed  CAS  Google Scholar 

  • Xu, R.H., Peck, R.M., Li, D.S., et al., Basic FGF and Suppression of BMP Signaling Sustain Undifferentiated Proliferation of Human ES Cells, Nat. Methods, 2005, vol. 2, pp. 185–190.

    Article  PubMed  CAS  Google Scholar 

  • Ying, Q.L., Nichols, J., Chambers, I., and Smith, A., BMP Induction of Id Proteins Suppresses Differentiation and Sustains Embryonic Stem Cell Self-Renewal in Collaboration with STAT3, Cell, 2003, vol. 115, pp. 281–292.

    Article  PubMed  CAS  Google Scholar 

  • Ying, Q., Wray, J., Nichols, J., et al., The Ground State of Embryonic Stem Cell Self-Renewal, Nature, 2008, vol. 453, pp. 519–523.

    Article  PubMed  CAS  Google Scholar 

  • Yoo, S.J., Yoon, B.S., Kim, J.M., et al., Efficient Culture System for Human Embryonic Stem Cells Using Autologous Human Embryonic Stem Cell-Derived Feeder Cells, Exp. Mol. Med., 2005, vol. 37, pp. 399–407.

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Kol’tsov Institute of Developmental Biology, Russian Academy of Sciences, Moscow, 119334, Russia

    N. V. Lifantseva & O. F. Gordeeva

  2. Institute of Cytology, Russian Academy of Sciences, St. Petersburg, 194064, Russia

    A. M. Koltsova & G. G. Poljanskaya

Authors
  1. N. V. Lifantseva
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. A. M. Koltsova
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. G. G. Poljanskaya
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. O. F. Gordeeva
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to O. F. Gordeeva.

Additional information

Original Russian Text © N.V. Lifantseva, A.M. Koltsova, G.G. Poljanskaya, O.F. Gordeeva, 2013, published in Ontogenez, 2013, Vol. 44, No. 1, pp. 10–23.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Lifantseva, N.V., Koltsova, A.M., Poljanskaya, G.G. et al. Expression of TGFβ family factors and FGF2 in mouse and human embryonic stem cells maintained in different culture systems. Russ J Dev Biol 44, 7–18 (2013). https://doi.org/10.1134/S1062360413010050

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1134/S1062360413010050

Keywords

Search

Navigation