Neural patterning of P19 cells derived embryoid bodies: an analogic perspective

Abstract

Abstract Research conducted on neuron differentiation from different cells is very limited. This research examined the patterning of differentiated neural cells in embryoid bodies (EBs) of two groups, hanging drop cells and aggregate cells obtained from P19 cells. P19 cells are a type of mouse embryonic carcinoma cells that can irreversibly differentiate into nerve cells by retinoic acid. P19 cell line was cultured in Minimum Essential Medium Alpha Medium in two hanging drop and aggregation methods. After 4 days, the EBs obtained from both methods were suspended into control and 1μM RA groups. A Real-Time PCR method and an inverted microscope were applied to evaluate neurons 8 days after induction. Using both hanging drop and aggregation methods, long and branched processes emerged from different parts of the cells by the end of the eighth day. Evaluation and comparison of the expression level of neural differentiation markers, including Nes, TUB βIII, and Map2, showed no significant difference between the two groups. However, aggregation group neurons exhibited higher levels of Nes mRNA. Neuronal identity markers, which represent the identity of the resulting neurons in EBs, revealed no significant difference between hanging drop and aggregation groups. The two above-mentioned methods can result in significant and similar differentiation of EBs into motor neuron-like neurons with spatial identity along the rostro-caudal axis under RA induction. This can be considered as a basis for nerve cell differentiation and transplantation in vitro and in vivo.