Skip to main content
SpringerLink
Log in

Optimization of Differentiation of Nonhuman Primate Pluripotent Cells Using a Combinatorial Approach

  • Protocol
  • First Online:
Neural Stem Cells

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1919))

  • 1725 Accesses

Abstract

The directed differentiation of pluripotent stem cells to a desired lineage often involves complex and lengthy protocols. In order to study the requirements for differentiation in a systematic way, we present here methodology for an iterative approach using combinations of small molecules and biological factors. The factors are used in a cyclical process in which the best combination of factors and concentrations is selected in one round of testing, followed by a modification of the combination and subsequent rounds. While this may produce the desired differentiation in the cell population under study, it is also possible that other strategies may be needed to optimize the differentiation process. These strategies are described in this chapter.

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 129.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Hardcover Book
USD 169.99
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. Farnsworth SL, Qiu Z, Mishra A, Hornsby PJ (2013) Directed neural differentiation of induced pluripotent stem cells from non-human primates. Exp Biol Med 238:276–284

    Article  CAS  Google Scholar 

  2. Tsutsui H, Valamehr B, Hindoyan A, Qiao R, Ding X, Guo S, Witte ON, Liu X, Ho CM, Wu H (2011) An optimized small molecule inhibitor cocktail supports long-term maintenance of human embryonic stem cells. Nat Commun 2:167

    Article  CAS  Google Scholar 

  3. Wu Y, Zhang Y, Mishra A, Tardif SD, Hornsby PJ (2010) Generation of induced pluripotent stem cells from newborn marmoset skin fibroblasts. Stem Cell Res 4:180–188

    Article  CAS  Google Scholar 

  4. Mishra A, Qiu Z, Farnsworth SL, Hemmi JJ, Li M, Pickering AV, Hornsby PJ (2016) Induced pluripotent stem cells from nonhuman primates. Methods Mol Biol 1357:183–193

    Article  CAS  Google Scholar 

  5. Hong CC, Yu PB (2009) Applications of small molecule BMP inhibitors in physiology and disease. Cytokine Growth Factor Rev 20:409–418

    Article  CAS  Google Scholar 

  6. Hollnagel A, Oehlmann V, Heymer J, Ruther U, Nordheim A (1999) Id genes are direct targets of bone morphogenetic protein induction in embryonic stem cells. J Biol Chem 274:19838–19845

    Article  CAS  Google Scholar 

  7. Ying QL, Nichols J, Chambers I, Smith A (2003) BMP induction of id proteins suppresses differentiation and sustains embryonic stem cell self-renewal in collaboration with STAT3. Cell 115:281–292

    Article  CAS  Google Scholar 

  8. Inman GJ, Nicolas FJ, Callahan JF, Harling JD, Gaster LM, Reith AD, Laping NJ, Hill CS (2002) SB-431542 is a potent and specific inhibitor of transforming growth factor-beta superfamily type I activin receptor-like kinase (ALK) receptors ALK4, ALK5, and ALK7. Mol Pharmacol 62:65–74

    Article  CAS  Google Scholar 

  9. Farberov S, Meidan R (2016) Thrombospondin-1 affects bovine luteal function via transforming growth factor-Beta1-dependent and independent actions. Biol Reprod 94:25

    Article  Google Scholar 

  10. Gudey SK, Sundar R, Heldin CH, Bergh A, Landstrom M (2017) Pro-invasive properties of Snail1 are regulated by sumoylation in response to TGFbeta stimulation in cancer. Oncotarget 8:97703–97726

    Article  Google Scholar 

  11. De Jaime-Soguero A, Aulicino F, Ertaylan G, Griego A, Cerrato A, Tallam A, Del Sol A, Cosma MP, Lluis F (2017) Wnt/Tcf1 pathway restricts embryonic stem cell cycle through activation of the Ink4/Arf locus. PLoS Genet 13:e1006682

    Article  CAS  Google Scholar 

  12. Kang Y, Hodges A, Ong E, Roberts W, Piermarocchi C, Paternostro G (2014) Identification of drug combinations containing imatinib for treatment of BCR-ABL+ leukemias. PLoS One 9:e102221

    Article  Google Scholar 

  13. Ding X, Matsuo K, Xu L, Yang J, Zheng L (2015) Optimized combinations of bortezomib, camptothecin, and doxorubicin show increased efficacy and reduced toxicity in treating oral cancer. Anti-Cancer Drugs 26:547–554

    Article  CAS  Google Scholar 

  14. Weiss A, Ding X, van Beijnum JR, Wong I, Wong TJ, Berndsen RH, Dormond O, Dallinga M, Shen L, Schlingemann RO, Pili R, Ho CM, Dyson PJ, van den Bergh H, Griffioen AW, Nowak-Sliwinska P (2015) Rapid optimization of drug combinations for the optimal angiostatic treatment of cancer. Angiogenesis 18:233–244

    Article  CAS  Google Scholar 

  15. Weiss A, Berndsen RH, Ding X, Ho CM, Dyson PJ, van den Bergh H, Griffioen AW, Nowak-Sliwinska P (2015) A streamlined search technology for identification of synergistic drug combinations. Sci Rep 5:14508

    Article  CAS  Google Scholar 

  16. Liu Q, Zhang C, Ding X, Deng H, Zhang D, Cui W, Xu H, Wang Y, Xu W, Lv L, Zhang H, He Y, Wu Q, Szyf M, Ho CM, Zhu J (2015) Preclinical optimization of a broad-spectrum anti-bladder cancer tri-drug regimen via the feedback system control (FSC) platform. Sci Rep 5:11464

    Article  Google Scholar 

  17. Ding X, Njus Z, Kong T, Su W, Ho CM, Pandey S (2017) Effective drug combination for Caenorhabditis elegans nematodes discovered by output-driven feedback system control technique. Sci Adv 3:eaao1254

    Article  CAS  Google Scholar 

  18. Chambers SM, Fasano CA, Papapetrou EP, Tomishima M, Sadelain M, Studer L (2009) Highly efficient neural conversion of human ES and iPS cells by dual inhibition of SMAD signaling. Nat Biotechnol 27:275–280

    Article  CAS  Google Scholar 

  19. Eiraku M, Watanabe K, Matsuo-Takasaki M, Kawada M, Yonemura S, Matsumura M, Wataya T, Nishiyama A, Muguruma K, Sasai Y (2008) Self-organized formation of polarized cortical tissues from ESCs and its active manipulation by extrinsic signals. Cell Stem Cell 3:519–532

    Article  CAS  Google Scholar 

  20. Zhou J, Su P, Li D, Tsang S, Duan E, Wang F (2010) High-efficiency induction of neural conversion in human ESCs and human induced pluripotent stem cells with a single chemical inhibitor of transforming growth factor beta superfamily receptors. Stem Cells 28:1741–1750

    Article  CAS  Google Scholar 

  21. Morizane A, Doi D, Kikuchi T, Nishimura K, Takahashi J (2011) Small-molecule inhibitors of bone morphogenic protein and activin/nodal signals promote highly efficient neural induction from human pluripotent stem cells. J Neurosci Res 89:117–126

    Article  CAS  Google Scholar 

  22. Yu PB, Hong CC, Sachidanandan C, Babitt JL, Deng DY, Hoyng SA, Lin HY, Bloch KD, Peterson RT (2008) Dorsomorphin inhibits BMP signals required for embryogenesis and iron metabolism. Nat Chem Biol 4:33–41

    Article  CAS  Google Scholar 

  23. Vogt J, Traynor R, Sapkota GP (2011) The specificities of small molecule inhibitors of the TGF and BMP pathways. Cell Signal 23:1831–1842

    Article  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Cellular and Integrative Physiology and Barshop Institute for Longevity and Aging Studies, University of Texas Health San Antonio, San Antonio, TX, USA

    Steven L. Farnsworth, Zhifang Qiu, Anuja Mishra & Peter J. Hornsby

Authors
  1. Steven L. Farnsworth
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Zhifang Qiu
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Anuja Mishra
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Peter J. Hornsby
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Peter J. Hornsby .

Editor information

Editors and Affiliations

  1. Southwest National Primate Research Center, Texas Biomedical Research Institute, San Antonio, TX, USA

    Marcel M. Daadi

Rights and permissions

Reprints and permissions

Copyright information

© 2019 Springer Science+Business Media, LLC, part of Springer Nature

About this protocol

Check for updates. Verify currency and authenticity via CrossMark

Cite this protocol

Farnsworth, S.L., Qiu, Z., Mishra, A., Hornsby, P.J. (2019). Optimization of Differentiation of Nonhuman Primate Pluripotent Cells Using a Combinatorial Approach. In: Daadi, M. (eds) Neural Stem Cells. Methods in Molecular Biology, vol 1919. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-9007-8_14

Download citation

  • DOI: https://doi.org/10.1007/978-1-4939-9007-8_14

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-9005-4

  • Online ISBN: 978-1-4939-9007-8

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation