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Neural Differentiation of Human Embryonic Stem Cells at the Ultrastructural Level

Published online by Cambridge University Press:  18 January 2010

Jennifer L. Mumaw ,
Dave Machacek ,
John P. Shields ,
Mahesh C. Dodla ,
Sujoy K. Dhara  and
Steve L. Stice
Jennifer L. Mumaw
Affiliation:
University of Georgia, Regenerative Bioscience Center, Rhodes Center for Animal and Dairy Sciences, 425 River Road, Room 427, Athens, GA 30602-2771, USA
Dave Machacek
Affiliation:
ArunA Biomedical, Rhodes Center for Animal and Dairy Sciences, 425 River Road, Room 452, Athens, GA 30602-2771, USA
John P. Shields
Affiliation:
University of Georgia, Center for Ultrastructural Research, 151 Barrow Hall, Athens, GA 30602-2771, USA
Mahesh C. Dodla
Affiliation:
University of Georgia, Regenerative Bioscience Center, Rhodes Center for Animal and Dairy Sciences, 425 River Road, Room 427, Athens, GA 30602-2771, USA
Sujoy K. Dhara
Affiliation:
University of Georgia, Regenerative Bioscience Center, Rhodes Center for Animal and Dairy Sciences, 425 River Road, Room 427, Athens, GA 30602-2771, USA
Steve L. Stice*
Affiliation:
University of Georgia, Regenerative Bioscience Center, Rhodes Center for Animal and Dairy Sciences, 425 River Road, Room 427, Athens, GA 30602-2771, USA
*
Corresponding author. E-mail: sstice@uga.edu
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Abstract

Neurodegerative disorders affect millions of people worldwide. Neural cells derived from human embryonic stem cells (hESC) have the potential for cell therapies and/or compound screening for treating affected individuals. While both protein and gene expression indicative of a neural phenotype has been exhibited in these differentiated cells, ultrastuctural studies thus far have been lacking. The objective of this study was to correlate hESC to neural differentiation culture conditions with ultrastructural changes observed in the treated cells. We demonstrate here that in basic culture conditions without growth factors or serum we obtain neural morphology. The addition of brain-derived neurotrophic factor (BDNF) and serum to cultures resulted in more robust neural differentiation. In addition to providing cues such as cell survival or lineage specification, additional factors also altered the intracellular structures and cell morphologies. Even though the addition of BDNF and serum did not increase synaptic formation, altered cellular structures such as abundant polyribosomes and more developed endoplasmic reticulum indicate a potential increase in protein production.

Keywords

embryonic stem cellsultrastructureneuralhumanneural differentiationdevelopment
Type
Biological Imaging: Techniques Development and Applications
Information
Microscopy and Microanalysis , Volume 16 , Issue 1 , February 2010 , pp. 80 - 90
Copyright
Copyright © Microscopy Society of America 2010

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References

REFERENCES

Baharvand, H., Jafary, H., Massumi, M. & Ashtiani, S.K. (2006). Generation of insulin-secreting cells from human embryonic stem cells. Dev Growth Differ 48(5), 323–332.CrossRefGoogle ScholarPubMed
Blanco, V., Lopez Camelo, J. & Carri, N.G. (2001). Growth inhibition, morphological differentiation and stimulation of survival in neuronal cell type (Neuro-2a) treated with trophic molecules. Cell Biol Int 25(9), 909–917.Google Scholar
Cho, Y.M., Kwon, S., Pak, Y.K., Seol, H.W., Choi, Y.M., Park Do, J., Park, K.S. & Lee, H.K. (2006). Dynamic changes in mitochondrial biogenesis and antioxidant enzymes during the spontaneous differentiation of human embryonic stem cells. Biochem Biophys Res Commun 348(4), 1472–1478.CrossRefGoogle ScholarPubMed
Condic, M.L. & Letourneau, P.C. (1997). Ligand-induced changes in integrin expression regulate neuronal adhesion and neurite outgrowth. Nature 389(6653), 852–856.Google Scholar
Deitch, J.S. & Banker, G.A. (1993). An electron microscopic analysis of hippocampal neurons developing in culture: Early stages in the emergence of polarity. J Neurosci 13(10), 4301–4315.Google Scholar
Erickson, C.A. & Trinkaus, J.P. (1976). Microvilli and blebs as sources of reserve surface membrane during cell spreading. Exp Cell Res 99(2), 375–384.Google Scholar
Halloran, M.C. & Berndt, J.D. (2003). Current progress in neural crest cell motility and migration and future prospects for the zebrafish model system. Dev Dyn 228(3), 497–513.CrossRefGoogle ScholarPubMed
Heo, J.S., Lee, M.Y. & Han, H.J. (2007). Sonic hedgehog stimulates mouse embryonic stem cell proliferation by cooperation of Ca2+/protein kinase C and epidermal growth factor receptor as well as Gli1 activation. Stem Cells 25(12), 3069–3080.CrossRefGoogle Scholar
Huettner, J.E., Lu, A., Qu, Y., Wu, Y., Kim, M. & McDonald, J.W. (2006). Gap junctions and connexon hemichannels in human embryonic stem cells. Stem Cells 24(7), 1654–1667.CrossRefGoogle ScholarPubMed
Hurst, J.H., Mumaw, J., Machacek, D.W., Sturkie, C., Callihan, P., Stice, S.L. & Hooks, S.B. (2008). Human neural progenitors express functional lysophospholipid receptors that regulate cell growth and morphology. BMC Neurosci 9, 118.CrossRefGoogle ScholarPubMed
Hurst, P.R., Jefferies, K., Eckstein, P. & Wheeler, A.G. (1978). An ultrastructural study of preimplantation uterine embryos of the rhesus monkey. J Anat 126(Pt 1), 209–220.Google Scholar
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(2), 88–95.CrossRefGoogle ScholarPubMed
Johkura, K., Cui, L., Asanuma, K., Okouchi, Y., Ogiwara, N. & Sasaki, K. (2004). Cytochemical and ultrastructural characterization of growing colonies of human embryonic stem cells. J Anat 205(4), 247–255.Google Scholar
Johnson, M.A., Weick, J.P., Pearce, R.A. & Zhang, S.C. (2007). Functional neural development from human embryonic stem cells: Accelerated synaptic activity via astrocyte coculture. J Neurosci 27(12), 3069–3077.CrossRefGoogle ScholarPubMed
Karasek, M., Swiltoslawski, J. & Zieliniska, A. (2004). Ultrastructure of the central nervous system: The basics. Folia Neuropathol 42(Suppl B), 1–9.Google Scholar
Lange, K. (2002). Role of microvillar cell surfaces in the regulation of glucose uptake and organization of energy metabolism. Am J Physiol Cell Physiol 282(1), C1–C26.CrossRefGoogle ScholarPubMed
Leahy, A., Xiong, J.W., Kuhnert, F. & Stuhlmann, H. (1999). Use of developmental marker genes to define temporal and spatial patterns of differentiation during embryoid body formation. J Exp Zool 284(1), 67–81.3.0.CO;2-O>CrossRefGoogle ScholarPubMed
Luft, J.H. (1961). Improvements in epoxy resin embedding methods. J Biophys Biochem Cytol 9, 409–414.CrossRefGoogle ScholarPubMed
Lyser, K.M. (1968). Early differentiation of motor neuroblasts in the chick embryo as studied by electron microscopy. II. Microtubules and neurofilaments. Dev Biol 17(2), 117–142.CrossRefGoogle ScholarPubMed
Maddox-Hyttel, P., Alexopoulos, N.I., Vajta, G., Lewis, I., Rogers, P., Cann, L., Callesen, H., Tveden-Nyborg, P. & Trounson, A. (2003). Immunohistochemical and ultrastructural characterization of the initial post-hatching development of bovine embryos. Reproduction 125(4), 607–623.Google Scholar
Meller, K. (1979). Scanning electron microscope studies on the development of the nervous system in vivo and in vitro. Int Rev Cytol 56, 23–56.Google Scholar
Nikas, G. (1999). Cell-surface morphological events relevant to human implantation. Hum Reprod 14(S2), 37–44.CrossRefGoogle ScholarPubMed
Oh, S.K., Kim, H.S., Ahn, H.J., Seol, H.W., Kim, Y.Y., Park, Y.B., Yoon, C.J., Kim, D.W., Kim, S.H. & Moon, S.Y. (2005). Derivation and characterization of new human embryonic stem cell lines: SNUhES1, SNUhES2, and SNUhES3. Stem Cells 23(2), 211–219.Google Scholar
Park, S.H., Kook, M.C., Kim, E.Y., Park, S. & Lim, J.H. (2004). Ultrastructure of human embryonic stem cells and spontaneous and retinoic acid-induced differentiating cells. Ultrastruct Pathol 28(4), 229–238.CrossRefGoogle ScholarPubMed
Sathananthan, A.H. & Trounson, A. (2005). Human embryonic stem cells and their spontaneous differentiation. Ital J Anat Embryol 110(2-S1), 151–157.Google Scholar
Shin, S., Dalton, S. & Stice, S.L. (2005). Human motor neuron differentiation from human embryonic stem cells. Stem Cells Dev 14(3), 266–269.CrossRefGoogle ScholarPubMed
Shin, S., Mitalipova, M., Noggle, S., Tibbitts, D., Venable, A., Rao, R. & Stice, S.L. (2006). Long-term proliferation of human embryonic stem cell-derived neuroepithelial cells using defined adherent culture conditions. Stem Cells 24(1), 125–138.Google Scholar
Van Blerkom, J. & Motta, P.M. (1989). Ultrastructure of Human Gametogenesis and Early Embryogenesis. Boston, MA: Kluwer Academic Publishers.CrossRefGoogle Scholar
Waterman, R.E. (1979). Scanning electron microscope studies of central nervous system development. Birth Defects Orig Artic Ser 15(3), 55–77.Google ScholarPubMed