Skip to main content
SpringerLink
Log in

Three-Dimensional Culture for Expansion and Differentiation of Embryonic Stem Cells

  • Protocol
  • First Online:
Book cover Human Embryonic and Induced Pluripotent Stem Cells

Part of the book series: Springer Protocols Handbooks ((SPH))

Abstract

Although embryonic stem (ES) cells have the regenerative capability of producing any tissue in our body, the development of efficient methods to generate a large number of ES cells with high purity has still remained a technical challenge before they can be used routinely in clinical trials. In this chapter, we describe a simple procedure to establish an ES cell 3D culture system through alginate-poly-l-lysine-alginate (APA) microencapsulation. The APA microcapsule provides an environment that promotes better cell growth and differentiation without requiring feeder layers.

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

Access this chapter

Log in via an institution
Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 169.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 219.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 299.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Odorico, J.S., Kaufman, D.S., Thomson, J.A. (2001) Multilineage Differentiation from Human Embryonic Stem Cell Lines. Stem Cells 19, 193–204.

    Article  PubMed  CAS  Google Scholar 

  2. Itskovitz-Eldor, J., Schuldiner, M., Karsenti, D., Eden, A., Yanuka, O., Amit, M., Soreq, H., Benvenisty, N. (2000) Differentiation of human embryonic stem cells into embryoid bodies compromising the three embryonic germ layers. Mol Med 6, 88.

    Google Scholar 

  3. Keller, G.M. (1995) In vitro differentiation of embryonic stem cells. Curr Opin Cell Biol 7, 862–869.

    Article  PubMed  CAS  Google Scholar 

  4. Xu, R.-H., Peck, R.M., Li, D.S., Feng, X., Ludwig, T., Thomson, J.A. (2005) Basic FGF and suppression of BMP signaling sustain undifferentiated proliferation of human ES cells. Nat Meth 2, 185–190.

    Article  CAS  Google Scholar 

  5. Bigdeli, N., Andersson, M., Strehl, R., Emanuelsson, K., Kilmare, E., Hyllner, J., Lindahl, A. (2008) Adaptation of human embryonic stem cells to feeder-free and matrix-free culture conditions directly on plastic surfaces. J Biotechnol 133, 146–153.

    Article  PubMed  CAS  Google Scholar 

  6. Gerecht, S., Burdick, J., Ferreira, L., Townsend, S., Langer, R., Vunjak-Novakovic, G. (2007) Hyaluronic acid hydrogel for controlled ­self-renewal and differentiation of human ­embryonic stem cells. Proc Natl Acad Sci 104, 11298.

    Article  PubMed  CAS  Google Scholar 

  7. Chen, H.F., Kuo, H.C., Chien, C.L., Shun, C.T., Yao, Y.L., Ip, P.L., Chuang, C.Y., Wang, C.C., Yang, Y.S., Ho, H.N. (2007) Derivation, characterization and differentiation of human embryonic stem cells: comparing serum-containing versus serum-free media and evidence of germ cell differentiation. Hum Reprod 22, 567–577.

    Article  PubMed  Google Scholar 

  8. Wang, L., Schulz, T.C., Sherrer, E.S., Dauphin, D.S., Shin, S., Nelson, A.M., Ware, C.B., Zhan, M., Song, C.-Z., Chen, X., Brimble, S.N., McLean, A., Galeano, M.J., Uhl, E.W., D’Amour, K.A., Chesnut, J.D., Rao, M.S., Blau, C.A., Robins, A.J. (2007) Self-renewal of human embryonic stem cells requires insulin-like growth factor-1 receptor and ERBB2 receptor signaling. Blood 110, 4111–4119.

    Article  PubMed  CAS  Google Scholar 

  9. Ellerström, C., Strehl, R., Noaksson, K., Hyllner, J., Semb, H. (2007) Facilitated Expansion of Human Embryonic Stem Cells by Single-Cell Enzymatic Dissociation. Stem Cells 25, 1690–1696.

    Article  PubMed  Google Scholar 

  10. Mallon, B.S., Park, K.-Y., Chen, K.G., Hamilton, R.S., McKay, R.D.G. (2006) Toward xeno-free culture of human embryonic stem cells. Int J Biochem Cell Biol 38, 1063–1075.

    Article  PubMed  CAS  Google Scholar 

  11. Skottman, H., Hovatta, O. (2006) Culture conditions for human embryonic stem cells. Reproduction 132, 691–698.

    Article  PubMed  CAS  Google Scholar 

  12. Hassell, T., Gleave, S., Butler, M. (1991) Growth inhibition in animal cell culture. Appl Biochem Biotechnol 30, 29–41.

    Article  PubMed  CAS  Google Scholar 

  13. Ferreira, L.S., Gerecht, S., Fuller, J., Shieh, H.F., Vunjak-Novakovic, G., Langer, R. (2007) Bioactive hydrogel scaffolds for controllable vascular differentiation of human embryonic stem cells. Biomaterials 28, 2706–2717.

    Article  PubMed  CAS  Google Scholar 

  14. Tielens, S., Declercq, H., Gorski, T., Lippens, E., Schacht, E., Cornelissen, M. (2007) Gelatin-Based Microcarriers as Embryonic Stem Cell Delivery System in Bone Tissue Engineering:  An in-Vitro Study. Biomacromolecules 8, 825–832.

    Article  PubMed  CAS  Google Scholar 

  15. Nie, Y., Bergendahl, V., Hei, D.J., Jones, J.M., Palecek, S.P. (2009) Scalable culture and cryopreservation of human embryonic stem cells on microcarriers. Biotechnol Prog 25, 20–31.

    Article  PubMed  CAS  Google Scholar 

  16. Hwang, Y.S., Cho, J., Tay, F., Heng, J.Y.Y., Ho, R., Kazarian, S.G., Williams, D.R., Boccaccini, A.R., Polak, J.M., Mantalaris, A. (2009) The use of murine embryonic stem cells, alginate encapsulation, and rotary microgravity bioreactor in bone tissue engineering. Biomaterials 30, 499–507.

    Article  PubMed  CAS  Google Scholar 

  17. Lock, L.T., Tzanakakis, E.S. (2009) Expansion and Differentiation of Human Embryonic Stem Cells to Endoderm Progeny in a Microcarrier Stirred-Suspension Culture. Tissue Eng Part A 15, 2051–2063.

    Article  PubMed  CAS  Google Scholar 

  18. Levenberg, S., Huang, N., Lavik, E., Rogers, A., Itskovitz-Eldor, J., Langer, R. (2003) Differentiation of human embryonic stem cells on three-dimensional polymer scaffolds. Proc Natl Acad Sci USA 100, 12741.

    Article  PubMed  CAS  Google Scholar 

  19. Mao, G.-h., Chen, G.-a., Bai, H.-y., Song, T.-r., Wang, Y.-x. (2009) The reversal of hyperglycaemia in diabetic mice using PLGA scaffolds seeded with islet-like cells derived from human embryonic stem cells. Biomaterials 30, 1706–1714.

    Article  PubMed  CAS  Google Scholar 

  20. Gerecht-Nir, S., Cohen, S., Ziskind, A., Itskovitz-Eldor, J. (2004) Three-dimensional porous alginate scaffolds provide a conducive environment for generation of well-vascularized embryoid bodies from human embryonic stem cells. Biotechnol Bioeng 88, 313–320.

    Article  PubMed  CAS  Google Scholar 

  21. Wang, X., Wang, W., Ma, J., Guo, X., Yu, X., Ma, X. (2006) Proliferation and Differentiation of Mouse Embryonic Stem Cells in APA Microcapsule: A Model for Studying the Interaction between Stem Cells and Their Niche. Biotechnol Prog 22, 791–800.

    Article  PubMed  CAS  Google Scholar 

  22. Maguire, T., Novik, E., Schloss, R., Yarmush, M. (2006) Alginate-PLL microencapsulation: Effect on the differentiation of embryonic stem cells into hepatocytes. Biotechnol Bioeng 93, 581–591.

    Article  PubMed  CAS  Google Scholar 

  23. Dang, S.M., Gerecht-Nir, S., Chen, J., Itskovitz-Eldor, J., Zandstra, P.W. (2004) Controlled, Scalable Embryonic Stem Cell Differentiation Culture. Stem Cells 22, 275–282.

    Article  PubMed  Google Scholar 

  24. Dean, S.K., Yulyana, Y., Williams, G., Sidhu, K.S., Tuch, B.E. (2006) Differentiation of Encapsulated Embryonic Stem Cells After Transplantation. Transplantation 82, 1175–1184.

    Article  PubMed  Google Scholar 

  25. Siti-Ismail, N., Bishop, A.E., Polak, J.M., Mantalaris, A. (2008) The benefit of human embryonic stem cell encapsulation for prolonged feeder-free maintenance. Biomaterials 29, 3946–3952.

    Article  PubMed  CAS  Google Scholar 

  26. Batorsky, A., Liao, J., Lund, A.W., Plopper, G.E., Stegemann, J.P. (2005) Encapsulation of adult human mesenchymal stem cells within collagen-agarose microenvironments. Biotechnol Bioeng 92, 492–500.

    Article  PubMed  CAS  Google Scholar 

  27. Li, X., Liu, T., Song, K., Yao, L., Ge, D., Bao, C., Ma, X., Cui, Z. (2006) Culture of Neural Stem Cells in Calcium Alginate Beads. Biotechnol Prog 22, 1683–1689.

    PubMed  CAS  Google Scholar 

  28. Ma, X., Vacek, I., Sun, A. (1994) Generation of alginate-poly-l-lysine-alginate (APA) biomicrocapsules: the relationship between the membrane strength and the reaction conditions. Artif Cells Blood Substit Biotechnol 22, 43–69.

    Article  CAS  Google Scholar 

  29. Peirone, M., Ross, C.J.D., Hortelano, G., Brash, J.L., Chang, P.L. (1998) Encapsulation of various recombinant mammalian cell types in different alginate microcapsules. J Biomed Mater Res 42, 587–596.

    Article  PubMed  CAS  Google Scholar 

  30. Orive, G., Hernández, R., Gascón, A., Calafiore, R., Chang, T., De Vos, P., Hortelano, G., Hunkeler, D., Lacík, I., Shapiro, A., Pedraz, J. (2003) Cell encapsulation: promise and ­progress. Nat Med 9, 104–107.

    Article  PubMed  CAS  Google Scholar 

  31. Li, H., Jiang, H., Wang, C., Duan, C., Ye, Y., Su, X., Kong, Q., Wu, J., Guo, X. (2006) Comparison of two types of alginate microcapsules on stability and biocompatibility in vitro and in vivo. Biomedical Materials 1, 42.

    Article  PubMed  CAS  Google Scholar 

  32. Lim, F., Sun, A.M. (1980) Microencapsulated islets as bioartificial endocrine pancreas. Science 210, 908–910.

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Laboratory of Biomedical Material Engineering, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian, PR China

    Guang-wei Sun, Xiao-xi Xu, Nan Li, Ying Zhang & Xiao-jun Ma

  2. Graduate School of the Chinese Academy, Becjing, PR China

    Xiao-xi Xu

Authors
  1. Guang-wei Sun
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Xiao-xi Xu
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Nan Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Ying Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Xiao-jun Ma
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Guang-wei Sun .

Editor information

Editors and Affiliations

  1. , College of Engineering, University of Arkansas, University of Arkansas 1, Fayetteville, 72701, Arkansas, USA

    Kaiming Ye

  2. , College of Engineering, University of Arkansas, University of Arkansas 1, Fayetteville, 72701, Arkansas, USA

    Sha Jin

Rights and permissions

Reprints and permissions

Copyright information

© 2011 Humana Press

About this protocol

Cite this protocol

Sun, Gw., Xu, Xx., Li, N., Zhang, Y., Ma, Xj. (2011). Three-Dimensional Culture for Expansion and Differentiation of Embryonic Stem Cells. In: Ye, K., Jin, S. (eds) Human Embryonic and Induced Pluripotent Stem Cells. Springer Protocols Handbooks. Humana Press. https://doi.org/10.1007/978-1-61779-267-0_5

Download citation

  • DOI: https://doi.org/10.1007/978-1-61779-267-0_5

  • Published:

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-61779-266-3

  • Online ISBN: 978-1-61779-267-0

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation