Investigating neural differentiation of mouse P19 embryonic stem cells in a time-dependent manner by bioinformatic, microscopic and transcriptional analyses

Background As an available cell line, mouse pluripotent P19 has been widely employed for neuronal differentiation studies. In this research, by applying the in vitro differentiation of this cell line into neuron-like cells through retinoic acid (RA) treatment, the roles of some genes including DNMT3B, ICAM1, IRX3, JAK2, LHX1, SOX9, TBX3 and THY1 in neural differentiation was investigated.Methods and results Bioinformatics, microscopic, and transcriptional studies were conducted in a time-dependent manner after RA-induced neural differentiation. According to bioinformatics studies, we determined the engagement of the metabolic and developmental super-pathways and pathways in neural cell differentiation, particularly focusing on the considered genes. According to our qRT-PCR analyses, JAK2, SOX9, TBX3, LHX1 and IRX3 genes were found to be significantly over expressed in a time-dependent manner (p < 0.05). In addition, the significant downregulation of THY1, DNMT3B and ICAM1 genes was observed during the experiment (p < 0.05). The optical microscopic investigation showed that the specialized extensions of the neuron-like cells were revealed on day 8 after RA treatment.Conclusion Accordingly, the neural differentiation of P19 cell line and the role of the considered genes during the differentiation were proved. However, our results warrant further in vivo studies.

Automated summary

In this study, the researchers investigated the neural differentiation of the mouse pluripotent P19 cell line and the roles of several genes during the differentiation process, including DNMT3B, ICAM1, IRX3, JAK2, LHX1, SOX9, TBX3, and THY1. Through bioinformatics, microscopic, and transcriptional studies, the researchers identified the involvement of metabolic and developmental super-pathways and pathways in neural cell differentiation. qRT-PCR analyses revealed significant upregulation of JAK2, SOX9, TBX3, LHX1, and IRX3 genes, and downregulation of THY1, DNMT3B, and ICAM1 genes during the experiment. Optical microscopic investigation showed the appearance of specialized extensions of neuron-like cells on day 8 after retinoic acid treatment. Further in vivo studies are needed to validate these results.