Derivation of phenotypically diverse neural culture from hESC by combining adherent and dissociation methods

Journal of Neuroscience Methods
Ye LiuDavid W Howells

Abstract

Differentiation of human embryonic stem cells (hESCs) into distinct neural lineages has been widely studied. However, preparation of mixed yet neurochemically mature populations, for the study of neurological diseases involving mixed cell types has received less attention. We combined two commonly used differentiation methods to provide robust and reproducible cultures in which a mixture of primarily GABAergic and Glutamatergic neurons was obtained. Detailed characterisation by immunocytochemistry (ICC) and quantitative real-time PCR (qPCR) assessed the neurochemical phenotype, and the maturation state of these neurons. We found that once neurospheres (NSs) had attached to the culture plates, proliferation of neural stem cell was suppressed. Neuronal differentiation and synaptic development then occurred after 21 days in vitro (DIV). By 49DIV, there were large numbers of neurochemically and structurally mature neurons. The qPCR studies indicated that expression of GABAergic genes increased the most (93.3-fold increase), followed by glutamatergic (51-fold increase), along with smaller changes in expression of cholinergic (3-fold increase) and dopaminergic genes (6-fold increase), as well as a small change in glial cell marker ex...Continue Reading

References

Nov 6, 1998·Science·J A ThomsonJ M Jones
Jan 5, 1999·Journal of Neuroscience Methods·C N SvendsenM A Caldwell
Mar 11, 1999·Current Opinion in Neurobiology·M Hynes, A Rosenthal
Nov 22, 2001·Experimental Neurology·M K CarpenterM S Rao
Dec 4, 2001·Nature Biotechnology·B E ReubinoffT Ben-Hur
Sep 21, 2004·Nature Reviews. Neuroscience·Henry MarkramCaizhi Wu
Aug 6, 2005·Molecular and Cellular Neurosciences·Pavel ItsyksonBenjamin E Reubinoff
May 25, 2007·The Journal of Neuroscience : the Official Journal of the Society for Neuroscience·Jakob von EngelhardtHannah Monyer
Jan 8, 2008·Differentiation; Research in Biological Diversity·Sujoy K DharaSteven L Stice
Mar 29, 2008·Methods in Molecular Biology·Mirella Dottori, Martin F Pera
Jun 13, 2008·Nature Protocols·Thomas D Schmittgen, Kenneth J Livak
Sep 1, 2009·Journal of Neuroscience Methods·Mathilda Zetterström AxellMaurice Curtis
Nov 13, 2009·Methods in Molecular Biology·Xiao-Qing Zhang, Su-Chun Zhang
May 23, 2012·Journal of Tissue Engineering and Regenerative Medicine·Laura Ylä-OutinenSusanna Narkilahti
Aug 15, 2014·The Journal of Neuroscience : the Official Journal of the Society for Neuroscience·Lydie MorelYongjie Yang
May 31, 2016·Stem Cells International·Piya PrajumwongsParinya Noisa

❮ Previous
Next ❯

Related Concepts

Related Feeds

Cell Migration

Cell migration is involved in a variety of physiological and pathological processes such as embryonic development, cancer metastasis, blood vessel formation and remoulding, tissue regeneration, immune surveillance and inflammation. Here is the latest research.