Abstract

The P19CL6 mouse embryonic carcinoma cells efficiently differentiate into cardiac muscle cells in the presence of DMSO. A reporter plasmid for cardiac muscle differentiation was constructed by connecting the CMV enhancer and a 250 bp MLC-2v promoter in front of the GFP gene to further evaluate the role of the CMV enhancer. This plasmid (pCBVenh/MLC-2vpro/EGFP) was stably introduced into P19CL6 cells, and the transfectant differentiated into cardiomyocytes with DMSO. Upon DMSO addition, GFP was immediately transcribed (within 2 days) and the amount of the transcript increased with cultivation. Concomitantly, GFP fluorescence was detected in the cells under a microscope. However, native MLC-2v was transcribed later on day 4. This expression time course is different from that of GFP. Clearly the CMV enhancer responded immediately to DMSO. Since GATA DNA-binding proteins play crucial roles in the initiation of cardiomyocyte differentiation, such a response could be ascribed to the presence of multiple GATA motifs in the enhancer sequence but not in the native MLC-2v promoter. Thus the CMV enhancer may be not only useful for gene therapy and monitoring cell differentiation but also the study of the role of GATA transcription factors expressed in P19CL6 cells.

Highlights

  • P19CL6 cells derived from P19 embryonic carcinoma cells can efficiently differentiate into cardiac muscle cells in the presence of 1% dimethyl sulfoxide (DMSO) [1]

  • The clone was cultured in the presence and absence of 1% DMSO, and green fluorescence derived from green fluorescent protein (GFP) was monitored under a microscope

  • P19CL6 cells are used as a model of differentiation of cardiac myoblasts into myocytes [1] [8]

Read more

Summary

Introduction

P19CL6 cells derived from P19 embryonic carcinoma cells can efficiently differentiate into cardiac muscle cells in the presence of 1% dimethyl sulfoxide (DMSO) [1]. Such a property is a good tool for the study of cardiac myocyte differentiation in vitro. The GATA-4 and the MEF2C and Tbx transcription factors play crucial roles in cardiac myocyte differentiation [4]. Genetic analysis suggested that NKX2.5 and GATA-6 together with these three transcription factors are essential for cardiac development [5] [6]

Methods
Results
Conclusion
Full Text
Paper version not known

Talk to us

Join us for a 30 min session where you can share your feedback and ask us any queries you have

Schedule a call

Disclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.