Affiliation:
1. Islamic Azad University of Tabriz
2. Iran University of Medical Sciences
3. Ardabil University of Medical Sciences
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.
Publisher
Research Square Platform LLC
Reference32 articles.
1. Neurogenesis Using P19 Embryonal Carcinoma Cells;Au - Leszczyński P;JoVE,2019
2. Characterization of P19 cells during retinoic acid induced differentiation;Babuška V;Prague Med Rep,2010
3. From embryonal carcinoma cells to neurons: the P19 pathway;Bain G;BioEssays: news and reviews in molecular cellular and developmental biology,1994
4. An essential role for retinoid signaling in anteroposterior neural specification and neuronal differentiation;Blumberg B;Semin Cell Dev Biol,1997
5. Developmental regulation of the Hox genes during axial morphogenesis in the mouse;Deschamps J;Development,2005