Paracetamol perturbs neuronal arborization and disrupts the cytoskeletal proteins SPTBN1 and TUBB3 in both human and chicken in vitro models

ANDA: an open-source tool for automated image analysis of in vitro neuronal cells

“…2 ), as well as freshly plated and differentiated PC12Ns (Additional file 1 : Figure S2), differentiated NT2Ns (Additional file 1 : Figure S3), neuroblastoma SH-SY5Y (Additional file 1 : Figure S4) and in vitro day 8 E16 primary mouse neurons (Additional file 1 : Figure S5). Details of the cultivation of these five cell types are described in Additional file information [ 18 , 36 , 37 ]. To obtain phase contrast images of the cells we used the live-cell imaging platforms IncuCyte ® ZOOM for the CGN and PC12N cells, and IncuCyte® S3 for the NT2N, SH-SY5Y and primary mouse neuronal cells (Additional file methods).…”

“…6 ). AraC is a mitotic inhibitor often used as an inhibitor of glial cell proliferation in neuronal cell cultures [ 18 ]. AraC treatment is therefore expected to impact number of cell bodies over time.…”

“…We have previously shown that ANDA is able to quantify the impact CGN and NT2N neurite lengths and neurite attachment points upon treatment with increasing doses of the analgesic paracetamol (acetaminophen) [ 18 ]. After 72 h, lower doses of paracetamol had no significant effect on NT2Ns, whereas the highest dose showed an effect on neurite length and neurite attachment points.…”

ANDA: An open-source tool for automated image analysis of neuronal differentiation

“…The quality of ANDA's ability to quantify neuronal differentiation metrics was assessed using outlines of identified structures in freshly plated and in vitro day 3 CGN cells (Figure 2), as well as freshly plated and differentiated PC12N (Supplementary Figure 2) and NT2N (Supplementary Figure 3). Details of the cultivation of these three cell types are described in supplementary information [18,33]. To obtain phase contrast images of the cells we used the live-cell imaging platforms IncuCyte® ZOOM for the CGN and PC12N cells, and IncuCyte® S3 for the NT2N cells (Supplementary methods).…”

“…When microscopy is applied in these studies, the focus has been on changes to different morphologic parameters of neuronal cells, often in a high-throughput manner [16][17][18][19][20][21][22][23]. The use of label-free time-course phase contrast microscopy has increased our understanding on the rise, development, and maturation of neuronal networks without being confounded by factors such as phototoxicity [23][24][25].…”