title: In vitro modelling of cortical neurogenesis by sequential induction of human umbilical cord blood stem cells.
abstract: Neurogenesis of excitatory neurons in the developing human cerebral neocortex is a complex and dynamic set of processes and the exact mechanisms controlling the specification of human neocortical neuron subtypes are poorly understood due to lack of relevant cell models available. It has been shown that the transcription factors Pax6, Tbr2 and Tbr1, which are sequentially expressed in the rodent neocortex, regulate and define corticogenesis of glutamatergic neocortical neurons. In humans the homologues of these genes are generally expressed in a similar pattern, but with some differences. In this study, we used purified human umbilical cord blood stem cells, expressing pluripotency marker genes (OCT4, SOX2 and NANOG), to model human neocortical neurogenesis in vitro. We analyzed the expression patterns of PAX6, TBR2 and TBR1, at both protein and mRNA levels, throughout the 24 days of a sequential neuronal induction protocol. Their expression patterns correlated with those found in the developing human neocortex where they define different developmental stages of neocortical neurons. The derived cord blood neuron-like cells expressed a number of neuronal markers. They also expressed components of glutamatergic neurotransmission including glutamate receptor subunits and transporters, and generated calcium influxes upon stimulation with glutamate. Thus we have demonstrated that it is possible to model neocortical neurogenesis using cord blood stem cells in vitro. This may allow detailed analysis of the molecular mechanisms regulating neocortical neuronal specification, thus aiding the development of potential therapeutic tools for diseases and injuries of the cerebral cortex.
authors.affiliation: Institute of Genetic Medicine, International Centre for Life, Newcastle University, Newcastle-upon-Tyne, NE1 3BZ, United Kingdom.
authors.forename: Hamad
authors.initials: H
authors.lastname: Ali
authors.forename: Nicolas
authors.initials: N
authors.lastname: Forraz
authors.forename: Colin P
authors.initials: CP
authors.lastname: McGuckin
authors.forename: Marcin
authors.initials: M
authors.lastname: Jurga
authors.forename: Susan
authors.initials: S
authors.lastname: Lindsay
authors.forename: Bui Kar
authors.initials: BK
authors.lastname: Ip
authors.forename: Andrew
authors.initials: A
authors.lastname: Trevelyan
authors.forename: Christina
authors.initials: C
authors.lastname: Basford
authors.forename: Saba
authors.initials: S
authors.lastname: Habibollah
authors.forename: Sajjad
authors.initials: S
authors.lastname: Ahmad
authors.forename: Gavin J
authors.initials: GJ
authors.lastname: Clowry
authors.forename: Nadhim
authors.initials: N
authors.lastname: Bayatti
mesh_terms: D000943:Antigens, Differentiation / Q000235:genetics / Q000378:metabolism; D020013:Calcium Signaling; D002478:Cells, Cultured; D005136:Eye Proteins / Q000235:genetics / Q000378:metabolism; D005312:Fetal Blood / Q000166:cytology*; D005333:Fetus / Q000166:cytology; D020869:Gene Expression Profiling; D018507:Gene Expression Regulation, Developmental; D018698:Glutamic Acid / Q000502:physiology; D018398:Homeodomain Proteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D008954:Models, Biological; D000071317:Nanog Homeobox Protein; D019579:Neocortex / Q000166:cytology*; D055495:Neurogenesis; D009474:Neurons / Q000378:metabolism; D050814:Octamer Transcription Factor-3 / Q000235:genetics / Q000378:metabolism; D000071841:PAX6 Transcription Factor; D051761:Paired Box Transcription Factors / Q000235:genetics / Q000378:metabolism; D016271:Proto-Oncogene Proteins c-myc / Q000235:genetics / Q000378:metabolism; D016194:Receptors, N-Methyl-D-Aspartate / Q000378:metabolism; D012097:Repressor Proteins / Q000235:genetics / Q000378:metabolism; D055748:SOXB1 Transcription Factors / Q000235:genetics / Q000378:metabolism; D013234:Stem Cells / Q000378:metabolism / Q000502:physiology*; D020825:T-Box Domain Proteins / Q000235:genetics / Q000378:metabolism; D050598:Vesicular Glutamate Transport Protein 1 / Q000378:metabolism
chemical_list: D000943:Antigens, Differentiation; D005136:Eye Proteins; D018398:Homeodomain Proteins; C489427:MYC protein, human; C486780:NANOG protein, human; D000071317:Nanog Homeobox Protein; D050814:Octamer Transcription Factor-3; D000071841:PAX6 Transcription Factor; C000605730:PAX6 protein, human; C494173:POU5F1 protein, human; D051761:Paired Box Transcription Factors; D016271:Proto-Oncogene Proteins c-myc; D016194:Receptors, N-Methyl-D-Aspartate; D012097:Repressor Proteins; C493464:SLC17A7 protein, human; C527782:SOX2 protein, human; D055748:SOXB1 Transcription Factors; D020825:T-Box Domain Proteins; C574705:TBR1 protein, human; D050598:Vesicular Glutamate Transport Protein 1; D018698:Glutamic Acid