Directed differentiation of mouse P19 embryonal carcinoma cells to neural cells in a serum- and retinoic acid-free culture medium

Verma, Isha and Seshagiri, Polani B (2018) Directed differentiation of mouse P19 embryonal carcinoma cells to neural cells in a serum- and retinoic acid-free culture medium. In: IN VITRO CELLULAR & DEVELOPMENTAL BIOLOGY-ANIMAL, 54 (8). pp. 567-579.

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Abstract

P19 embryonal carcinoma cells (EC-cells) provide a simple and robust culture system for studying neural development. Most protocols developed so far for directing neural differentiation of P19 cells depend on the use of culture medium supplemented with retinoic acid (RA) and serum, which has an undefined composition. Hence, such protocols are not suitable for many molecular studies. In this study, we achieved neural differentiation of P19 cells in a serum- and RA-free culture medium by employing the knockout serum replacement (KSR) supplement. In the KSR-containing medium, P19 cells underwent predominant differentiation into neural lineage and by day 12 of culture, neural cells were present in 100% of P19-derived embryoid bodies (EBs). This was consistently accompanied by the increased expression of various neural lineage-associated markers during the course of differentiation. P19-derived neural cells comprised of NES+ neural progenitors (degrees similar to 46%), TUBB3(+) immature neurons (similar to 6%), MAP2(+) mature neurons (similar to 2%), and GFAP(+) astrocytes (similar to 50%). A heterogeneous neuronal population consisting of glutamatergic, GABAergic, serotonergic, and dopaminergic neurons was generated. Taken together, our study shows that the KSR medium is suitable for the differentiation of P19 cells to neural lineage without requiring additional (serum and RA) supplements. This stem cell differentiation system could be utilized for gaining mechanistic insights into neural differentiation and for identifying potential neuroactive compounds.

Item Type:	Journal Article
Publication:	IN VITRO CELLULAR & DEVELOPMENTAL BIOLOGY-ANIMAL
Publisher:	SPRINGER
Additional Information:	Copy right for this article belong to SPRINGER, 233 SPRING ST, NEW YORK, NY 10013 USA
Keywords:	Embryonal carcinoma cells; P19 cells; Differentiation; Embryoid body; Knockout serum replacement; Neural cells
Department/Centre:	Division of Biological Sciences > Molecular Reproduction, Development & Genetics
Date Deposited:	24 Sep 2018 15:37
Last Modified:	24 Sep 2018 15:37
URI:	http://eprints.iisc.ac.in/id/eprint/60693

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