Abstract
In this chapter, we describe an effective and reproducible protocol for neural differentiation of human pluripotent stem cells in three dimensional (3D) collagen and MartigelTM gels. We have used this protocol to generate embryoid bodies (EBs) from dissociated suspension cultures of human embryonic stem cells (hESCs) and induced pluripotent stem cells (hiPSCs) which are immobilized in 3D collagen or MartigelTM gels. The gel-entrapped EBs differentiated into robust Pax6+ neuroectodermal cells of neural rosettes in N2 neural-inducing medium, then into nestin+ neural progenitors and further differentiated into TuJ1+ neurons, GFAP+ astrocytes, and O4+ oligodendrocytes. This 3D neural differentiation model, derived from human ES and iPS cells, has potential applications in the study of early human embryonic development, drug screening, and cell therapy.
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References
Zhang SC, Wernig M, Duncan ID, Brüstle O, Thomson JA. (2001) In vitro differentiation of transplantable neural precursors from human embryonic stem cells. Nat Biotechnol. 19:1129–1133.
Ma W., Tavakoli T., Derby E., Serebryakova Y., Rao MS., Mattson MP. (2008a) Cell-extracellular matrix interactions regulate neural differentiation of human embryonic stem cells. BMC Developmental Biology, 8:90.
Hu BY, Zhang SC. (2009) Differentiation of spinal motor neurons from pluripotent human stem cells. Nat Protoc. 4:1295–304.
Tavakoli T., Derby E., Serebryakova Y., Rao MS., Mattson MP., Ma W. (2009) Self-Renewal and differentiation capabilities are variable between human embryonic stem cell lines I3, I6 and BG01V. BMC Cell Biology, 10:44.
Hwang NS, Varghese S, Elisseeff J. (2008) Controlled differentiation of stem cells. Adv Drug Deliv Rev. 60:199–214.
O’Connor SM., Stenger DA., Shaffer KM., Maric D., Barker JL, Ma W. (2000a) Primary neural precursor O cell expansion, differentiation and cytosolic Ca2+ response in three-dimentional collagen gel. Journal of Neuroscience Methods, 102:187–195.
O’Connor SM., Andreadis JD., Shaffer KM., Ma W., Pancrazio J., Stenger DA. (2000b) Immobilization of neural cells in three-dimensional matrices for biosensor applications. Biosensors & Bioelectronics, 14:871–881.
O’Connor SM., Stenger DA., Shaffer KM., Ma W. (2001) Survival and neurite outgrowth of rat cortical neurons in three-dimensional agarose and collagen gel matrices. Neuroscience Letter, 304:189–193.
O’Shaughnessy TJ., Lin HJ., Ma W. (2003) Functional synapse formation among rat cortical neurons grown on three-dimensional collagen gels. Neuroscience Letter, 340:169–172.
Lin JH., O’Shaughnessy TJ., Kelly J., Ma W. (2004) Neural stem and progenitor cell differentiation in a cell-collagen-bioreactors culture system. Developmental Brain Research, 153:163–173.
Ma W., Fitzgerald W., O’Shaughnessy TJ., Lin HJ., Liu QL., Maric D., Alkon DL., Barker JL. (2004) CNS stem and progenitor cell differentiation into functional neuronal circuits in three-dimensional collagen gels. Experimental Neurology, 190: 276–288.
Ma W., Chen S., Fitzgerald W., Maric D., Lin HJ., O’Shaughnessy TJ., Kelly J., Liu X-H., Barker JL. (2005) Three-dimensional collagen gel networks for neural stem cell-based neural tissue engineering. Macromolecular Symposia, 227:327–333.
Ma W., Tavakoli T, Chen S., Maric M., Lin HJ., O’Shaughnessy TJ., Barker JL. (2008b) Reconstruction of functional cortical-like tissues from neural stem and progenitor cells. Tissue Engineering, 10: 1673–1685.
Stenger DA., Gross GW., Keefer EW., Shaffer KM., Andreadis JD., Ma W., Pancrazio JJ. (2001) Detection of physiologically active compounds using cell-based biosensors. Trends in Biotechnology, 19:304–309.
Kunlin Jin, et al. (2010) Transplantation of Human Precursor Cells in Martigel Scaffolding Improves Outcome from Focal Cerebral Ischemia after Delayed Postischemic Treatment in Rats. J Cereb Blood Flow Metab. 30(3): 534–544.
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© 2011 Humana Press
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Derby, E., Yin, D., Gao, WQ., Ma, W. (2011). Neural Differentiation of Human ES and iPS Cells in Three-Dimensional Collagen and Martigelâ„¢ Gels. 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_32
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DOI: https://doi.org/10.1007/978-1-61779-267-0_32
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Online ISBN: 978-1-61779-267-0
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