Patients' stem cells differentiation in a 3D environment as a promising experimental tool for the study of Amyotrophic Lateral Sclerosis

The database includes the raw data of the article “Patients’ stem cells differentiation in a 3D environment as a promising experimental tool for the study of Amyotrophic Lateral Sclerosis”. Aim of this work was to demonstrate the validity of 3D cultures for the differentiation of stem cells for the study of Amyotrophic Lateral Sclerosis in a more realistic model.

The database contains the data obtained by the following evaluations: a) immunofluorescence analysis for the study of cells’ proliferation during differentiation b) confocal microscopy for the evaluation of differentiation markers c) RT-qPCR for the confirmation of stem cells’ differentiation d) immunofluorescence analysis for the evaluation of the electrophysiological characteristics of differentiated NSC34 cells.

We first reprogrammed healthy and sALS PBMCs in iPSCs and differentiated them in NSCs in 2D. NSCs were then included in Cellink Bioink and printed. We differentiated NSCs first in MNPs and finally in MNs and evaluated every step of 3D differentiation in comparison with 2D. We found that NSCs maintain a good viability and proliferation rate during the differentiation in 3D. Moreover, we confirmed the good differentiation by confocal microscopy and RT-qPCR by the use of specific markers for every specific differentiation step. We concluded that cell’s inclusion in the hydrogel does not interfere with the correct differentiation process. Moreover, Bioprinting allows the study of cells in a 3D environment which better mimics tissue organization. Furthermore, RT-qPCR results suggest that the 3D environment increases the differentiation of CTRL MNPs in mature MNs when compared to 2D.

Finally, we evidenced that the electrophysiological characteristics of cells are maintained after the inclusion and the printing.

Such evidence confirmed that 3D Bioprinting can be considered a good model for the study of ALS pathogenesis.