Skip to main content
SpringerLink
Log in

Deciphering the Importance of Three Key Media Components in Human Embryonic Stem Cell Cultures

  • Published:
Biotechnology Letters Aims and scope Submit manuscript

Abstract

Development of a serum free, feeder-free (SFFF) culture platform for human embryonic stem cells (hESC) will be important for the expansion of hESC for future cell therapy applications. However, currently, culture of hESC consists of a combination of basal media, basic fibroblast growth factor (bFGF), serum replacer (SR) and conditioned media (CM) from feeders, and it is unclear which components of the mixture are absolutely critical in the maintenance of hESC. To evaluate the relative contributions of these media components in the development of SFFF culture, each was systematically eliminated and pluripotency assayed by dual embryonic stem cell markers, Oct-4 and TRA-1-60. We concluded that SR was the most critical component in the platform, followed by bFGF and CM produced by feeders, where down-regulation of Oct-4 occurred after 2, 5 and 5 passages, respectively, upon their withdrawal from the complete media.

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

  • S Assady G Maor M Amit J Itskovitz-Eldor KL Skorecki M Tzukerman (2001) ArticleTitleInsulin production by human embryonic stem cells Diabetes 50 1691–1697 Occurrence Handle1:CAS:528:DC%2BD3MXlslKntbc%3D Occurrence Handle11473026

    CAS  PubMed  Google Scholar 

  • AB Choo J Padmanabhan AC Chin SKW Oh (2004) ArticleTitleExpansion of pluripotent human embryonic stem cells on human feeders Biotechnol. Bioeng. 88 321–331 Occurrence Handle10.1002/bit.20247 Occurrence Handle1:CAS:528:DC%2BD2cXpsF2murg%3D Occurrence Handle15486939

    Article  CAS  PubMed  Google Scholar 

  • Choo AB, Padmanabhan J, Fong WJ, Chin AC, Oh SKW (2005) Immortalized feeders for the scale up of human embryonic stem cells in feeder and feeder free conditions. J. Biotechnol. (E-pub. ahead of print)

  • Pebay A, Wong RCB, Pitson SM, Wolvetang E, Peh GSL, Filipczyk A, Koh KLL, Tellis I, Nguyen LTV, Pera MF (2005) Essential roles of sphingosine-1-phosphate and platelet-derived growth factor in the maintenance of human embryonic stem cells. Stem Cells (E-pub. ahead of print)

  • MF Pera B Reubinoff A Trounson (2000) ArticleTitleHuman embryonic stem cells J. Cell Sci. 113 IssueIDPt 1 5–10 Occurrence Handle1:CAS:528:DC%2BD3cXhtValu7g%3D Occurrence Handle10591620

    CAS  PubMed  Google Scholar 

  • Price P, Goldsborough M, Tilkins ML (1998) Embryonic stem cell serum replacer. PCT/US98/00467: WO 98/30679

  • BE Reubinoff P Itsykson T Turetsky MF Pera E Reinhartz A Itzik T Ben Hur (2001) ArticleTitleNeural progenitors from human embryonic stem cells Nat. Biotechnol. 19 1134–1140 Occurrence Handle10.1038/nbt1201-1134 Occurrence Handle1:CAS:528:DC%2BD3MXptV2rt70%3D Occurrence Handle11731782

    Article  CAS  PubMed  Google Scholar 

  • JA Thomson J Itskovitz-Eldor SS Shapiro MA Waknitz JJ Swiergiel VS Marshall JM Jones (1998) ArticleTitleEmbryonic stem cell lines derived from human blastocysts Science 282 1145–1147 Occurrence Handle10.1126/science.282.5391.1145 Occurrence Handle1:CAS:528:DyaK1cXntleisLg%3D Occurrence Handle9804556

    Article  CAS  PubMed  Google Scholar 

  • L Wang L Li P Menendez C Cerdan M Bhatia (2005a) ArticleTitleHuman embryonic stem cells maintained in the absence of mouse embryonic fibroblasts or conditioned media are capable of hematopoietic development Blood 105 4598–4603 Occurrence Handle1:CAS:528:DC%2BD2MXlsVWjtb0%3D

    CAS  Google Scholar 

  • G Wang H Zhang Y Zhao J Li J Cai P Wang S Meng J Feng C Miao M Ding D Li H Deng (2005b) ArticleTitleNoggin and bFGF cooperate to maintain the pluripotency of human embryonic stem cells in the absence of feeder layers Biochem. Biophys. Res. Commun. 330 934–942 Occurrence Handle1:CAS:528:DC%2BD2MXivV2jurg%3D

    CAS  Google Scholar 

  • C Xu MS Inokuma J Denham K Golds P Kundu JD Gold MK Carpenter (2001) ArticleTitleFeeder-free growth of undifferentiated human embryonic stem cells Nature Biotechnol. 19 971–974 Occurrence Handle1:CAS:528:DC%2BD3MXntl2msbY%3D

    CAS  Google Scholar 

  • C Xu SMCK Police (2002) ArticleTitleCharacterization and enrichment of cardiomyocytes derived from human embryonic stem cells Circ. Res. 91 501–508 Occurrence Handle10.1161/01.RES.0000035254.80718.91 Occurrence Handle1:CAS:528:DC%2BD38XotFehurw%3D Occurrence Handle12242268

    Article  CAS  PubMed  Google Scholar 

  • C Xu E Rosler J Jiang JS Lebkowski JD Gold C O’Sullivan K Delavan-Boorsma M Mok A Bronstein MK Carpenter (2005a) ArticleTitleBasic fibroblast growth factor supports undifferentiated human embryonic stem cell growth without conditioned medium Stem Cells 23 315–323 Occurrence Handle10.1634/stemcells.2004-0211 Occurrence Handle1:CAS:528:DC%2BD2MXjtFyjtb4%3D

    Article  CAS  Google Scholar 

  • RH Xu RM Peck DS Li X Feng T Ludwig JA Thomson (2005b) ArticleTitleBasic FGF and suppression of BMP signalling sustain undifferentiated proliferation of human ES cells Nat. Methods 2 185–190 Occurrence Handle10.1038/nmeth744 Occurrence Handle1:CAS:528:DC%2BD2MXisVGisrc%3D

    Article  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Stem Cell Group, Bioprocessing Technology Institute, 20 Biopolis Way, #06–01 Centros, 138668, Singapore, Singapore

    Vanessa Ding, Andre B. H. Choo & Steve K. W. Oh

Authors
  1. Vanessa Ding
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Andre B. H. Choo
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Steve K. W. Oh
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Steve K. W. Oh.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Ding, V., Choo, A.B.H. & Oh, S.K.W. Deciphering the Importance of Three Key Media Components in Human Embryonic Stem Cell Cultures. Biotechnol Lett 28, 491–495 (2006). https://doi.org/10.1007/s10529-006-0005-8

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10529-006-0005-8

Keywords

Search

Navigation