Skip to main content

Enhanced proliferation of bone marrow mesenchymal stem cells by co-culture with TM4 mouse Sertoli cells: involvement of the EGF/PI3K/AKT pathway

Molecular and Cellular Biochemistry volume 393, pages 155–164 (2014)Cite this article

Abstract

Bone marrow mesenchymal stem cells (BM-MSCs) are considered as a promising option in the field of regenerative medicine and tissue engineering. However, little is known about how TM4 mouse Sertoli cells, which are known to enhance stem cells proliferation in co-culture, may influence the proliferation of BM-MSCs and which signaling pathways are involved in. To address these questions, an in vitro transwell system was used. We found that TM4 cells could produce soluble factors which enhanced the growth of BM-MSCs without inhibiting the multipotency. Furthermore, cell cycle analysis showed that co-culture with the TM4 cells accelerated the progress of BM-MSCs from the G1 to the S phase. The expression of the phospho-akt, mdm2, as well as pho-CDC2, and cyclin D1 were markedly upregulated in co-cultured BM-MSCs. The observed promoting effect was significantly inhibited by the administration of the PI3K/AKT inhibitor, LY294002. Among the various growth factors produced by TM4 cells, the epithelial growth factor (EGF) stimulated the proliferation of the BM-MSCs more significantly compared with the other growth factors examined in this study. Neutralization of EGF via a blocking antibody significantly limited the promoting growth effect in BM-MSCs. These results suggest that TM4 cells provide a favorable in vitro environment for BM-MSCs growth and imply the involvement of the EGF/PI3K/AKT pathway.

This is a preview of subscription content, access via your institution.

Access options

Buy single article

Instant access to the full article PDF.

43,34 €

Price includes VAT (Finland)
Tax calculation will be finalised during checkout.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8

References

  1. Ballas CB, Zielske SP, Gerson SL (2002) Adult bone marrow stem cells for cell and gene therapies: implications for greater use. J Cell Biochem 85:20–28

    Article  Google Scholar 

  2. Zhang F, Hong Y, Liang W, Ren T, Jing S, Lin J (2012) Co-culture with Sertoli cells promotes proliferation and migration of umbilical cord mesenchymal stem cells. Biochem Biophys Res Commun 427:86–90

    CAS  PubMed  Article  Google Scholar 

  3. Tsutsumi S, Shimazu A, Miyazaki K et al (2001) Retention of multilineage differentiation potential of mesenchymal cells during proliferation in response to FGF. Biochem Biophys Res Commun 288:413–419

    CAS  PubMed  Article  Google Scholar 

  4. Liu C, Yu J, Yang Y et al (2013) Effect of 1 mT sinusoidal electromagnetic fields on proliferation and osteogenic differentiation of rat bone marrow mesenchymal stromal cells. Bioelectromagnetics 34:453–464

    PubMed  Article  Google Scholar 

  5. Kermani S, Karbalaie K, Madani SH et al (2008) Effect of lead on proliferation and neural differentiation of mouse bone marrow-mesenchymal stem cells. Toxicol In Vitro 22:995–1001

    CAS  PubMed  Article  Google Scholar 

  6. Strobel LA, Hild N, Mohn D, Stark WJ et al (2013) Novel strontium-doped bioactive glass nanoparticles enhance proliferation and osteogenic differentiation of human bone marrow stromal cells. J Nanopart Res 15:1780

    Article  Google Scholar 

  7. Cai B, Meng F, Zhu S et al (2010) Arsenic trioxide induces the apoptosis in bone marrow mesenchymal stem cells by intracellular calcium signal and caspase-3 pathways. Toxicol Lett 193:173–178

    CAS  PubMed  Article  Google Scholar 

  8. Skinner MK (1993) Secretion of growth factors and other regulatory factors. In: Russell LA, Griswold MD (eds) The Sertoli cell: clearwater. Cache River Press, St. Louis

    Google Scholar 

  9. Dufour JM, Rajotte RV, Kin T, Korbutt GS (2003) Immunoprotection of rat islet xenografts by cotransplantation with sertoli cells and a single injection of antilymphocyte serum. Transplantation 75:1594–1596

    PubMed  Article  Google Scholar 

  10. Fan P, He L, Pu D et al (2011) Testicular Sertoli cells influence the proliferation and immunogenicity of co-cultured endothelial cells. Biochem Biophys Res Commun 404:829–833

    CAS  PubMed  Article  Google Scholar 

  11. Othberg AI, Willing AE, Cameron DF et al (1998) Trophic effect of porcine Sertoli cells on rat and human ventral mesencephalic cells and hNT neurons in vitro. Cell Transplant 7:157–164

    CAS  PubMed  Article  Google Scholar 

  12. Huo Y, Qiu W, Pan Q, Yao Y, Xing K, Lou MF (2009) Reactive oxygen species (ROS) are essential mediators in epidermal growth factor (EGF)-stimulated corneal epithelial cell proliferation, adhesion, migration, and wound healing. Exp Eye Res 89:876–886

    CAS  PubMed  Article  Google Scholar 

  13. Ilowski M, Putz C, Weiss TS, Brand S, Jauch KW, Hengstler JG, Thasler WE (2010) Augmenter of liver regeneration causes different kinetics of ERK1/2 and Akt/PKB phosphorylation than EGF and induces hepatocyte proliferation in an EGF receptor independent and liver specific manner. Biochem Biophys Res Commun 39:915–920

    Article  Google Scholar 

  14. Salehinejad P, Alitheen NB, Mandegary A et al (2013) Effect of EGF and FGF on the expansion properties of human umbilical cord mesenchymal cells. In Vitro Cell Dev Biol Anim 49:515–523

    CAS  PubMed  Article  Google Scholar 

  15. Xu Z, Yan L, Ge Y et al (2012) Effect of the calcium sensing receptor on rat bone marrow-derived mesenchymal stem cell proliferation through the ERK1/2 pathway. Mol Biol Rep 39:7271–7279

    CAS  PubMed  Article  Google Scholar 

  16. Ryu JM, Lee MY, Yun SP, Han HJ (2010) High glucose regulates cyclin D1/E of human mesenchymal stem cells through TGF-β1 expression via Ca2+/PKC/MAPKs and PI3K/Akt/mTOR signal pathways. J Cell Physiol 224:59–70

    CAS  PubMed  Google Scholar 

  17. Fukao T, Koyasu S (2003) PI3K and negative regulation of TLR signaling. Trends Immunol 24:358–363

    CAS  PubMed  Article  Google Scholar 

  18. Kong D, Yamori T (2008) Phosphatidylinositol 3-kinase inhibitors: promising drug candidates for cancer therapy. Cancer Sci 99:1734–1740

    CAS  PubMed  Article  Google Scholar 

  19. Golden LH, Insogna KL (2004) The expanding role of PI3-kinase in bone. Bone 34:3–12

    CAS  PubMed  Article  Google Scholar 

  20. Xuan Nguyen TL, Choi JW, Lee SB et al (2006) Akt phosphorylation is essential for nuclear translocation and retention in NGF-stimulated PC12 cells. Biochem Biophys Res Commun 349:789–798

    PubMed  Article  Google Scholar 

  21. Zhou L, Kong J, Zhuang Y, Chu J, Zhang S, Guo M (2013) Ex vivo expansion of bone marrow mesenchymal stem cells using microcarrier beads in a stirred bioreactor. Biotechnol Bioprocess Eng 18:173–184

    CAS  Article  Google Scholar 

  22. Mo X, Guo S, Xie H et al (2009) Variations in the ratios of co-cultured mesenchymal stem cells and chondrocytes regulate the expression of cartilaginous and osseous phenotype in alginate constructs. Bone 45:42–51

    PubMed  Article  Google Scholar 

  23. Sun J, Zhu H, Liu C et al (2013) GDNF Up-Regulates c-Myc transcription via the PI3K/Akt pathway to promote dairy goat male germline stem cells (mGSC) proliferation. J Integr Agric 12:1054–1065

    Article  Google Scholar 

  24. Sun B, Feng M, Tian X, Lu X et al (2012) DL-3-n-Butylphthalide protects rat bone marrow stem cells against hydrogen peroxide-induced cell death through antioxidation and activation of PI3K-Akt pathway. Neurosci Lett 516:247–252

    CAS  PubMed  Article  Google Scholar 

  25. Tamama K, Fan YH, Griffith LG et al (2006) Epidermal growth factor as a candidate for ex vivo expansion of bone marrow-derived mesenchymal stem cells. Stem Cells 24:686–695

    CAS  PubMed  Article  Google Scholar 

  26. Tie L, Luo L (2005) Epidermal growth factor-induced proliferation requires down-regulation of Pax6 in corneal epithelial cells. J Biol Chem 280:12988–12995

    Article  Google Scholar 

Download references

Acknowledgments

It was supported by the National Basic Research Program of China (No. 2011CB200900), the National High Technology Research & Development Program of China (No. 2012AA021201 and 2014AA021705), the National Special Fund for State Key Laboratory of Bioreactor Engineering (No. 2060204), and the Fundamental Research Funds for the Central Universities of ECUST.

Conflict of interest

The authors declare that there are no conflicts of interest.

Author information

Affiliations

  1. State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, P.O. Box 329, 130 Meilong Rd, Shanghai, 200237, P.R. China

    Huan Tian, Meijin Guo, Yingping Zhuang, Ju Chu & Siliang Zhang

Authors
  1. Huan Tian
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Meijin Guo
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Yingping Zhuang
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Ju Chu
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Siliang Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Meijin Guo.

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Tian, H., Guo, M., Zhuang, Y. et al. Enhanced proliferation of bone marrow mesenchymal stem cells by co-culture with TM4 mouse Sertoli cells: involvement of the EGF/PI3K/AKT pathway. Mol Cell Biochem 393, 155–164 (2014). https://doi.org/10.1007/s11010-014-2055-x

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11010-014-2055-x

Keywords

  • Bone marrow mesenchymal stem cells
  • TM4 cells
  • Co-culture
  • Cell proliferation
  • EGF/PI3K/AKT pathway