Lisa Bachmann, Germany
Ruhr-University Bochum Biochemistry IIAuthor Of 1 Presentation
DIFFERENTIATION AND CHARACTERIZATION OF CEREBRAL ORGANOIDS FROM HUMAN IPS CELLS WITH A PSEN1 MUTATION VS. ISOGENIC CONTROLS
Abstract
Aims
Within this study we aimed to develop a new cerebral organoid model to study mechanisms underlying familial Alzheimer’s disease. Human induced pluripotent stem cells (hiPSC) with a PSEN1 mutation and the isogenic control cells should be analysed in respect to their potential to differentiate cerebral organoids.
Methods
The methods included stem cell culture, differentiation of embryonic bodies and cerebral organoids, the freezing, slicing, and the staining of the organoids. Differentiation of cerebral organoids was assessed using primary antibodies against PAX6, SOX2, OCT4, MAP2, betaTubulinIII, and FOXG1. Antibodies against human beta amyloid, phosphorylated Tau, and AICD were used to identify Alzheimer disease-relevant changes in the PSEN+/+, PSEN+/-, and PSEN-/- (wt) organoid slices. The stained and mounted samples were imaged with the Leica TCS SP8 confocal microscope.
Results
The PSEN1 cerebral organoids revealed normal differentiation including development of PAX6+ precursor cells and mature bTubIII+ neurons, which together established a cortical plate-like brain structure. Immunofluorescence analysis of sliced organoids revealed cell-type specific APP processing and expression of PSEN1.
Conclusions
Due to the organization of PSEN1 cerebral organoids mimicking the human brain and due to their human origin, they correspond to a promising model system to study Alzheimer’s disease relevant pathways, which are of relevance for familial as well as for sporadic AD. Robust expression of APP and PSEN1 also empower the model system for drug treatment screens and identification of reliable biomarkers.