A differential response to newt regeneration extract by C2C12 and primary mammalian muscle cells.

Sarah Kawesa,Jason Vanstone,Catherine Tsilfidis

doi:10.1186/s13395-015-0044-8

Sarah Kawesa, Jason Vanstone + Show 1 more

Open Access

https://doi.org/10.1186/s13395-015-0044-8

Copy DOI

Abstract

BackgroundDedifferentiation, a process whereby differentiated cells lose their specialized characteristics and revert to a less differentiated state, plays a key role in the regeneration process in urodele amphibians such as the red spotted newt, Notophthalmus viridescens. Dedifferentiation of fully mature tissues is generally absent in mammalian cells. Previous studies have shown that mouse C2C12 multinucleated myotubes treated with extract derived from regenerating newt forelimbs can re-enter the cell cycle, fragment into mononucleated cells, and proliferate. However, this response has been difficult to replicate.MethodsWe isolated extract from early newt forelimb regenerates and assessed its effects on differentiation of proliferating primary and C2C12 myoblasts. We also treated fully differentiated primary and C2C12 myotube cultures with extract and assessed cell cycle re-entry and myotube fragmentation.ResultsWe have confirmed the results obtained in C2C12 cells and expanded these studies to also examine the effects of newt regeneration extracts on primary muscle cells. Newt extract can block differentiation of both C2C12 and primary myoblasts. Once differentiation is induced, treatment with newt extract causes cell cycle re-entry and fragmentation of C2C12 myotubes. Downregulation of p21 and muscle-specific markers is also induced. Primary myotubes also fragment in response to extract treatment, and the fragmented cells remain viable for long periods of time in culture. However, unlike C2C12 cells, primary muscle cells do not re-enter the cell cycle in response to treatment with newt extracts.ConclusionsDedifferentiation of fully mature muscle occurs during regeneration in the newt forelimb to contribute cells to the regeneration process. Our study shows that extracts derived from regenerating newt forelimbs can induce dedifferentiation, cell cycle re-entry, and fragmentation of mouse C2C12 cells but can only induce fragmentation in primary muscle cells.Electronic supplementary materialThe online version of this article (doi:10.1186/s13395-015-0044-8) contains supplementary material, which is available to authorized users.

Highlights

Dedifferentiation, a process whereby differentiated cells lose their specialized characteristics and revert to a less differentiated state, plays a key role in the regeneration process in urodele amphibians such as the red spotted newt, Notophthalmus viridescens
There were no apparent effects on cell cycle in myoblasts that were treated with 1° newt extract at 0.3 mg/ml in comparison to the control
Semiconfluent myoblasts were plated with the same number of cells in 1 ml of growth medium (GM) either in the presence or absence of newt extract

Summary

Introduction

Dedifferentiation, a process whereby differentiated cells lose their specialized characteristics and revert to a less differentiated state, plays a key role in the regeneration process in urodele amphibians such as the red spotted newt, Notophthalmus viridescens. Previous studies have shown that mouse C2C12 multinucleated myotubes treated with extract derived from regenerating newt forelimbs can re-enter the cell cycle, fragment into mononucleated cells, and proliferate. This response has been difficult to replicate. It is believed that the dedifferentiation process is the key to regeneration [4, 5] Through this process, differentiated cells re-enter the cell cycle and revert back to a progenitor phenotype [6,7,8] that can proliferate and re-differentiate into the various tissues of the regenerated structure

Methods

Results

Discussion

Conclusion