Human iPS to neuron (wt) 1
|Series:||IN_VITRO DIFFERENTIATION SERIES|
|Species:||Human (Homo sapiens)|
|Link to TET:||FILE|
|Sample providers :||Christine Wells|
|Primary cells or cell line:||primary cells|
|Time span:||18 days|
|Number of time points:||4|
Human induced pluripotent stem cells can generate every cell type of the human body and under the appropriate conditions recapitulate central aspects of embryonic development in the dish. To interrogate the transcriptome changes associated with the earliest steps of human brain development as recapitulated with human pluripotent stem cells we generated footprint-free induced pluripotent stem cells (iPSC) from control and Down syndrome fibroblasts using episomal reprogramming  and were next stepwise differentiated these iPSc into neuro-ectodermal cells (day6), neural stem cells (day12) and early neuronal progenitors (day 18) using an established neuronal differentiation protocol .
For this study three sample donors were used, two 2 control iPSC (C11 iPSC derived from CRL2429 Newborn Male Caucasian fibroblasts and C32 iPSC derived from CRL1502 12wk gestation Female Black fibroblasts) and two iPSC clones from one Down Syndrome individual (C11 and C18 from an Unknown Male Caucasian). Three replicates of each iPSc line were subjected to neuronal differentiation as described [1,2] and harvested at day 0, 6, 12, 18 of differentiation for RNA extraction (Fig 1 below).
Q-PCR validation of key neuronal marker genes was performed and published . The data show upregulation of mRNA expression of the neuronal marker genes PAX6  , beta-III-tubulin , SOX1  , DCX , SOX9 , SOX2  and MASH1  (Fig 2C and D) and robust expression of PAX6, beta-III-Tubulin and MAP2  protein expression (Fig 2A). we further expect that as cells exit from pluripotency that they will display a downregulation of the pluripotency transcription factor Oct4  and the DNA methyl transferase DNMT3B .
Profiled time course samples
|13421-144D1||iPS differentiation to neuron, control donor C11-CRL2429||day00||rep1|
|13422-144D2||iPS differentiation to neuron, control donor C11-CRL2429||day06||rep1|
|13423-144D3||iPS differentiation to neuron, control donor C11-CRL2429||day12||rep1|
|13424-144D4||iPS differentiation to neuron, control donor C11-CRL2429||day18||rep1|
|13425-144D5||iPS differentiation to neuron, control donor C11-CRL2429||day00||rep2|
|13426-144D6||iPS differentiation to neuron, control donor C11-CRL2429||day06||rep2|
|13427-144D7||iPS differentiation to neuron, control donor C11-CRL2429||day12||rep2|
|13428-144D8||iPS differentiation to neuron, control donor C11-CRL2429||day18||rep2|
|13429-144D9||iPS differentiation to neuron, control donor C11-CRL2429||day00||rep3|
|13430-144E1||iPS differentiation to neuron, control donor C11-CRL2429||day06||rep3|
|13431-144E2||iPS differentiation to neuron, control donor C11-CRL2429||day12||rep3|
|13432-144E3||iPS differentiation to neuron, control donor C11-CRL2429||day18||rep3|