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Neural Differentiation of Human Adipose Tissue-Derived Stem Cells

Protocol
First Online:
Part of the Methods in Molecular Biology book series (MIMB, volume 702)

Abstract

While adult stem cells can be induced to transdifferentiate into multiple lineages of cells or tissues, their plasticity and utility for human therapy remains controversial. In this chapter, we describe methods for the transdifferentiation of human adipose tissue-derived stem cells (ASCs) along neural lineages using in vitro and in vivo systems. The in vitro neural differentiation of ASCs has been reported by several groups using serum-free cytokine induction, butylated hydroxyanisole (BHA) chemical induction, and neurosphere formation in combination with the cytokines, such as brain-derived neurotrophic factor (BDNF) and basic fibroblast growth factor (bFGF). For in vivo neurogenic induction, ASCs are treated with BDNF and bFGF to form neurospheres in vitro and then delivered directly to the brain. In this chapter, several detailed protocols for the effective neurogenic induction of ASCs in vitro and in vivo are described. The protocols described herein can be applied to further molecular and mechanistic studies of neurogenic induction and differentiation of ASCs. In addition, these methods can be useful for differentiating ASCs for therapeutic intervention in central nervous system disorders.

Key words

Adipose tissue-derived mesenchymal stem cells Transdifferentiation Neurogenesis Reprogramming Neurosphere 
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References

  1. 1.
    Kim JH, Lee MR, Kim JH, Jee MK, Kang SK. 2008. Selenium induces improvement of stem cell behaviors in human adipose-tissue stromal cells via SAPK/JNK and stemness acting signals. Stem Cells 26:2724–2734.PubMedCrossRefGoogle Scholar
  2. 2.
    Kang SK, Putnam LA, Ylostalo J, Popescu IR, Dufour J, Belousov A, Bunnell BA. 2004. Neurogenesis of Rhesus adipose stromal cells. J Cell Sci 15:4289–4299.CrossRefGoogle Scholar
  3. 3.
    Kang SK, Putnam L, Dufour J, Ylostalo J, Jung JS, Bunnell BA. 2004. Expression of telomerase extends the lifespan and enhances osteogenic differentiation of adipose tissue-derived stromal cells. Stem Cells 22:1356–1372.PubMedCrossRefGoogle Scholar
  4. 4.
    Kim JH, Jee MK, Lee SY, Han TH, Kim BS, Kang KS, Kang SK. 2009. Regulation of adipose tissue stromal cells behaviors by endogenic Oct4 expression control. PLoS One 4:e7166.PubMedCrossRefGoogle Scholar
  5. 5.
    Safford KM, Hicok KC, Safford SD, Halvorsen YD, Wilkison WO, Gimble JM, Rice HE. 2002. Neurogenic differentiation of murine and human adipose-derived stromal cells. Biochem Biophys Res Commun 294:371–379.PubMedCrossRefGoogle Scholar
  6. 6.
    Krampera M, Marconi S, Pasini A, Galie M, Rigotti G, Mosna F, Tinelli M, Lovato L, Anghileri E, Andreini A, Pizzolo G, Sbarbati A, Bonetti B. 2007. Induction of neural-like differentiation in human mesenchymal stem cells derived from bone marrow, fat, spleen and thymus. Bone 40:382–390.PubMedCrossRefGoogle Scholar
  7. 7.
    Tseng PY, Chen CJ, Sheu CC, Yu, CW, Huang YS. 2007. Spontaneous differentiation of adult rat marrow stromal cells in a long-term culture. J Vet Med Sci 69:95–102.PubMedCrossRefGoogle Scholar
  8. 8.
    Kang SK, Yeo JE, Kang KS, Phinney DG. 2007. Cytoplasmic extracts from adipose tissue stromal cells alleviates secondary damage by modulating apoptosis and promotes functional recovery following spinal cord injury. Brain Pathol. 17:263–275.PubMedCrossRefGoogle Scholar
  9. 9.
    Kang SK, Shin MJ, Jung JS, Kim YG, Kim CH. 2006. Autologous adipose tissue-derived stromal cells for treatment of spinal cord injury. Stem Cells Dev. 15:583–594.PubMedCrossRefGoogle Scholar
  10. 10.
    Rosenstein K. 2003. Liquidation of a pharmacy’s prescription drug inventory. Health Law Can 24:18–27.PubMedGoogle Scholar
  11. 11.
    Lennmyr F, Ata KA, Terent A. 1998. Expression of vascular endothelial growth factor (VEGF) and its receptors (Flt-1 and Flk-1) following permanent and transient occlusion of the middle cerebral artery in the rat. J Neuropathol Exp Neurol 57:874–882.PubMedCrossRefGoogle Scholar
  12. 12.
    Zhu Y, Jin K, Greenberg DA. 2003. Vascular endothelial growth factor promotes proliferation of cortical neuron precursors by regulating E2F expression. FASEB J 17:186–193.PubMedCrossRefGoogle Scholar
  13. 13.
    Grafi G. 2004. How cells dedifferentiate: a lesson from plants. Dev Biol 268:1–6.PubMedCrossRefGoogle Scholar
  14. 14.
    Yourey PA, Gohari S, Alderson RF. 2000. Vascular endothelial cell growth factors promote the in vitro development of rat photoreceptor cells. J Neurosci 20:6781–6788.PubMedGoogle Scholar
  15. 15.
    Grafi G, Ben-Meir H, Avivi Y, Moshe M, Dahan Y, Zemach A. 2007. Histone methylation controls telomerase-independent telomere lengthening in cells undergoing dedifferentiation. Dev Biol 306:838–846.PubMedCrossRefGoogle Scholar
  16. 16.
    Jee MK, Kim JH, Han YM, Jung SJ, Kang KS, Kim DW, Kang SK. 2010. DHP-derivative and low oxygen tension effectively induces human adipose stromal cell reprogramming. PLOS ONE 9:5(2):e9026.Google Scholar
  17. 17.
    Akimoto T, Nonaka T, Harashima K, Sakurai H, Ishikawa H, Mitsuhashi N. 2004. Radicicol potentiates heat-induced cell killing in a human esophageal cancer cell line: the Hsp90 chaperone complex as a new molecular target for enhancement of thermosensitivity. Int J Radiat Biol 80:483–492.PubMedCrossRefGoogle Scholar
  18. 18.
    Ng DC, Bogoyevitch MA. 2000. The mechanism of heat shock activation of ERK mitogen-activated protein kinases in the interleukin 3-dependent proB cell line BaF3. J Biol Chem 275:40856–40866.PubMedCrossRefGoogle Scholar
  19. 19.
    Qian DX, Zhang HT, Ma X, Jiang XD, Xu RX. 2009. Comparison of the Efficiencies of three neural induction protocols in human adipose stromal cells. Neurochem Res. 35(4):572–579.Google Scholar
  20. 20.
    Padovan CS, Jahn K, Birnbaum T, Reich P, Sostak P, Strupp M, Straube A. 2003. Expression of neuronal markers in differentiated marrow stromal cells and CD133+ stem-like cells. Cell Transplant 12:839–848.PubMedGoogle Scholar
  21. 21.
    Kim YJ, Kim HK, Cho HH, Bae YC, Suh KS, Jung JS. 2007. Direct comparison of human mesenchymal stem cells derived from adipose tissues and bone marrow in mediating neovascularization in response to vascular ischemia. Cell Physiol Biochem 20:867–876.PubMedCrossRefGoogle Scholar
  22. 22.
    Hermann A, Gastl R, Liebau S, Popa MO, Fiedler J, Boehm BO. 2004. Efficient generation of neural stem cell-like cells from adult human bone marrow stromal cells. J Cell Sci 1:4411–4422.CrossRefGoogle Scholar
  23. 23.
    Ashjian PH, Elbarbary AS, Edmonds B, DeUgarte D, Zhu M, Zuk PA, Lorenz HP, Benhaim P, Hedrick MH. 2003. In vitro differentiation of human processed lipoaspirate cells into early neural progenitors. Plast Reconstr Surg 111:1922–1931.PubMedCrossRefGoogle Scholar
  24. 24.
    Woodbury D, Reynolds K, Black IB. 2002. Adult bone marrow stromal stem cells express germLine, ectodermal, endodermal, and mesodermal genes prior to neurogenesis. J Neurosci Res 69:908–917.PubMedCrossRefGoogle Scholar
  25. 25.
    Zuk PA, Zhu M, Ashjian P, De Ugarte DA, Huang JI, Mizuno H, Alfonso ZC, Fraser JK, Benhaim P, Hedrick MH 2002. Human adipose tissue is a source of multipotent stem cells. Mol Biol Cell 13:4279–4295.PubMedCrossRefGoogle Scholar
  26. 26.
    Franco Lamberta AP, Fraga Zandonai A, Bonatto D, Cantarelli Machado D, Pegas Henriques JA. 2009. Differentiation of human adipose-derived adult stem cells into neuronal tissue: Does it work? Differentiation 77:221–228.CrossRefGoogle Scholar
  27. 27.
    Hofstetter CP, Schwarz EJ, Hess D, Widenfalk J, El Manira A, Prockop DJ, Olson L. 2002. Marrow stromal cells form guiding strands in the injured spinal cord and promote recovery. Proc Natl Acad Sci USA 99:2199–2204.PubMedCrossRefGoogle Scholar
  28. 28.
    Goodell MA. 2003. Stem-cell “plasticity”: befuddled by the muddle. Curr Opin Hematol 10:208–213.PubMedCrossRefGoogle Scholar
  29. 29.
    Wagers AJ, Weissman IL. 2004. Plasticity of adult stem cells. Cell 116:639–648.PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  1. 1.Division of Regenerative MedicineTulane National Primate Research CenterCovingtonUSA
  2. 2.Center for Stem Cell Research and Regenerative MedicineTulane University School of MedicineNew OrleansUSA
  3. 3.Tulane National Primate Research CenterCovingtonUSA
  4. 4.Adult Stem Cell Research Center, Laboratory of Stem Cell and Tumor Biology, Department of Veterinary Public HealthCollege of Veterinary MedicineSeoulKorea

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