Abstract
In the last decade, there is an increasing application of induced pluripotent stem cells (iPSCs) for disease modeling. The iPSC technology enables the study of patient-specific neuronal cell lines in vitro to evaluate dysfunction at the cellular level and identify the responsible genetic factors. This approach might be particularly valuable for filling the gap of knowledge at the cellular and molecular levels underlying the pathophysiology of various neurodevelopmental and/or psychiatric disorders, such as attention-deficit hyperactivity disorder (ADHD). However, the invasiveness of skin biopsy or blood withdrawal might represent a major impediment in such protected population. Using hair derived keratinocytes as starting somatic cells circumvents this problem as sample collections can be performed non-invasively. Here we describe an improved, convenient, standardized and effective method to culture and reprogram hair derived keratinocytes from three healthy controls and one ADHD patient into iPSCs, which in turn will be used to generate differentiated neuronal cells. All the cell types were maintained in highly defined, serum-free conditions and showed expression of the respective key marker genes, assessed by both immunocytochemistry and qRT-PCR. The described in vitro personalized neuronal model has its advantage in modeling neurodevelopmental trajectories since it can recapitulate key processes of brain development at the cellular and molecular level and is intended to be used as for example studying ADHD etiopathology.
Highlights
The molecular and cellular mechanisms involved in neurodevelopmental disorders like attentiondeficit hyperactivity disorder (ADHD) are often poorly understood, mainly due to inaccessibility to brain tissue and limitations of the available model systems.As it is the case in other neurodevelopmental disorders, investigating the etiopathology of attention-deficit hyperactivity disorder (ADHD) at the molecular level is limited by a multitude of factors
As it is the case in other neurodevelopmental disorders, investigating the etiopathology of ADHD at the molecular level is limited by a multitude of factors
Gene expression analysis revealed an upregulated expression of telomerase reverse transcriptase (TERT), NANOG, octamer-binding transcription factor 3/4 (OCT3/4), lin-28 homolog A (LIN28A) and a downregulation of the keratinocyte marker keratin 14 (KRT14), compared to keratinocyte cultures (Figure 2 and Supplementary Table S3 for detailed relative expression)
Summary
The molecular and cellular mechanisms involved in neurodevelopmental disorders like attentiondeficit hyperactivity disorder (ADHD) are often poorly understood, mainly due to inaccessibility to brain tissue and limitations of the available model systems. As it is the case in other neurodevelopmental disorders, investigating the etiopathology of ADHD at the molecular level is limited by a multitude of factors. Modified animal models have recreated some of the prevailing symptoms of ADHD, but none of these models managed to reflect the disorder faithfully enough to lead to significant clinical implication (Giros et al, 1996; Carvalho et al, 2016). There is an urgent need to better understand the pathophysiology of the disorder in order to develop more effective, practical and individually tailored pharmaceutical therapies and to discover new predictive biomarkers
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Free) 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call
