Advertisement

Isolation and Propagation of Primary Human and Rodent Embryonic Neural Progenitor Cells and Cortical Neurons

  • Armine Darbinyan
  • Rafal Kaminski
  • Martyn K. White
  • Nune Darbinian
  • Kamel Khalili
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1078)

Abstract

The research on human neural progenitor cells holds great potential for the understanding the molecular programs that control differentiation of cells of glial and neuronal lineages and pathogenetic mechanisms of neurological diseases. Stem cell technologies provide also opportunities for pharmaceutical industry to develop new approaches for regenerative medicine. Here we describe the protocol for isolation and maintenance of neural progenitor cells and cortical neurons using human fetal brain tissue. This protocol can be successfully adapted for preparation of rodent neural and oligodendrocyte progenitor cells. While several methods for isolation of neural and oligodendrocyte progenitors from rodent brain tissue have been described, including techniques which use gene transfer and magnetic resonance beads, few methods are focused on derivation of human oligodendrocyte progenitor cells. Development of human culture provides the most physiologically relevant system for investigation of mechanisms which regulate function of oligodendrocyte, specifically of human origin.

Key words

Human neural progenitor cells Neurosphere Oligodendrocyte progenitor cells Neuron Oligodendrocyte 

References

  1. 1.
    Espinosa-Jeffrey A, Wakeman DR, Kim SU, Snyder EY, de Vellis J (2009) Culture system for rodent and human oligodendrocyte specification, lineage progression, and maturation. Curr Protoc Stem Cell Biol Chapter 2: Unit 2D.4Google Scholar
  2. 2.
    Pollard SM, Conti L, Sun Y, Goffredo D, Smith A (2006) Adherent neural stem (NS) cells from fetal and adult forebrain. Cereb Cortex 16(Suppl 1):i112–i120PubMedCrossRefGoogle Scholar
  3. 3.
    McCarthy KD, de Vellis J (1980) Preparation of separate astroglial and oligodendroglial cell cultures from rat cerebral tissue. J Cell Biol 85: 890–902PubMedCrossRefGoogle Scholar
  4. 4.
    Vicario-Abejón C, Johe KK, Hazel TG, Collazo D, McKay RD (1995) Functions of basic fibroblast growth factor and neurotrophins in the differentiation of hippocampal neurons. Neuron 15:105–114PubMedCrossRefGoogle Scholar
  5. 5.
    Rowitch DH, Kriegstein AR (2010) Developmental genetics of vertebrate glial-cell specification. Nature 468:214–222PubMedCrossRefGoogle Scholar
  6. 6.
    Darbinyan A, Kaminski R, White MK, Darbinian-Sarkissian N, Khalili K (2013) Polyomavirus JC infection inhibits differentiation of oligodendrocyte progenitor cells. J Neurosci Res 91(1):116–127. doi: 10.1002/jnr.23135 PubMedGoogle Scholar
  7. 7.
    Avellana-Adalid V (1996) Expansion of rat oligodendrocyte progenitors into proliferative "oligospheres" that retain differentiation potential. J Neurosci Res 45:558–570PubMedCrossRefGoogle Scholar
  8. 8.
    Neman J, de Vellis J (2012) A method for deriving homogenous population of oligodendrocytes from mouse embryonic stem cells. Dev Neurobiol 72(6):777–788. doi: 10.1002/dneu.22008 PubMedCrossRefGoogle Scholar
  9. 9.
    Merabova N, Kaminski R, Krynska B, Amini S, Khalili K, Darbinyan A (2012) JCV agnoprotein-induced reduction in CXCL5/LIX secretion by oligodendrocytes is associated with activation of apoptotic signaling in neurons. J Cell Physiol 227:3119–3127PubMedCrossRefGoogle Scholar
  10. 10.
    Merabova N, Kaniowska D, Kaminski R, Deshmane SL, White MK, Amini S, Darbinyan A, Khalili K (2008) JC virus agnoprotein inhibits in vitro differentiation of oligodendrocytes and promotes apoptosis. J Virol 82: 1558–1569PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2013

Authors and Affiliations

  • Armine Darbinyan
    • 1
  • Rafal Kaminski
    • 2
  • Martyn K. White
    • 2
  • Nune Darbinian
    • 2
  • Kamel Khalili
    • 2
  1. 1.Department of PathologyThe Mount Sinai Medical CenterNew YorkUSA
  2. 2.Department of NeuroscienceTemple University School of MedicinePhiladelphiaUSA

Personalised recommendations