Successful enrichment of cardiac progenitors following differentiation of EGFP-expressing transgenic mouse ES cells.

Sridharan, D and Seshagiri, P B (2017) Successful enrichment of cardiac progenitors following differentiation of EGFP-expressing transgenic mouse ES cells. In: ASCB/EMBO Meeting, DEC 02-06, 2017, Philadelphia, PA.

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Abstract

Derivation of embryonic stem (ES)-cell lines from genetically non-permissive mouse strains, such as FVB/N, has been difficult, despite this strain offering advantages for mouse transgenesis for developmental studies. We earlier generated β-actin promoter-driven enhanced green fluorescent protein (EGFP)-transgenic FVB/N mice, expressing EGFP in all cells. Here, by optimizing culture system and using RESGROTM ES-cell culture medium, we successfully derived EGFP-transgenic ES-cell line, ‘GS-2’ line, from F1 hybrid blastocysts, from wild-type 129/SvJ female X EGFPtransgenic homozygous FVB/N male. The GS-2 ES-cell line exhibited all defining criteria of a typical ES-cell line, including normal colony morphology and karyotype (40,XY), high replication-expansion efficiency (passages: >100), expression of pluripotent markers (Oct-4, Nanog, Sox-2, SSEA-1 and others) and, embryoid body (EB) development and EB differentiation to ecto-/meso-/endo-dermal cell types, expressing nestin, BMP-4 and α-fetoprotein, respectively. GS-2 ES-cells formed (i) teratoma containing three germ lineage-derived cell types, (ii) chimeric blastocysts and fetuses, following their aggregation with wild-type 8-cell embryos, (iii) functional cardiac clusters and (iv) predominantly neural cell types when EBs were developed in KOSR-supplemented medium. Taken together, we derived a robust EGFP-transgenic GS-2 ES-cell line, from a non-permissive transgenic (FVB/N) mouse by a single cross to 129/ SvJ wild-type mouse. The GS-2 ES-cell line exhibited full differentiation potential, in vitro/

Item Type:	Conference Proceedings
Publication:	MOLECULAR BIOLOGY OF THE CELL
Publisher:	AMER SOC CELL BIOLOGY, 8120 WOODMONT AVE, STE 750, BETHESDA, MD 20814-2755 USA
Additional Information:	Copy right for the article belong to AMER SOC CELL BIOLOGY, 8120 WOODMONT AVE, STE 750, BETHESDA, MD 20814-2755 USA
Department/Centre:	Division of Biological Sciences > Molecular Biophysics Unit
Date Deposited:	28 Mar 2018 16:18
Last Modified:	28 Mar 2018 16:18
URI:	http://eprints.iisc.ac.in/id/eprint/59414

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