Marco Thurner and Martin Deutsch from Innovacell Biotechnology AG, Austria, the Lonza CytoSMART Lab for muscle regeneration

Marco Thurner and Martin Deutsch, Innovacell Biotechnologie AG, Austria

 

 

 

 

 

In Vitro Differentiation of Skeletal Muscle Derived Cells

Research Interest

Innovacell Biotechnologie AG is an innovative company that concentrates on developing cell therapies for the treatment of incontinence. The concept of using patients’ own skeletal muscle derived cells (SMDCs) for the regeneration of defective sphincter muscles has been investigated by Innovacell in clinical trials [1, 2]. In order to quantify the myogenic potency of the SMDCs, the R&D department of Innovacell (headed by Dr. Rainer Marksteiner) has developed and patented an in vitro potency assay [3]. This assay is based on the principle that satellite cells isolated from the skeletal muscle [4] give rise to proliferating single nucleated cells that are able to fuse with each other to form multinucleated myotubes [5]. During this process myotubes express Acetylcholinesterase (AChE) in significant amounts, and the measurable AChE activity increases over time, which can be quantified in an accurate, rapid and simple way employing Ellman’s method [6]. AChE has a direct role in the regulation of muscle function that involves the hydrolysis of neurotransmitter Acetylcholine.


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The Experiment

To visualize the dynamic process of SMDC differentiation and myotube formation used in the described potency assay, biologists at the R&D department of Innovacell have used Lonza’s CytoSMART™ System. Human skeletal muscle derived cells were induced to differentiate for up to 7 days in vitro to form multinucleated myotubes in vitro. In parallel, AChE activity of the differentiating cells was measured according to the above-mentioned in vitro potency assay.

Image of myotubes taken with the CytoSMART System
 Graph showing AChE activity during the differentiation of SMDCs to multinucleated myotubes

 

Video of SMDCs differentiation into multinucleated myobtubes (recorded with the CytoSMART™ System for 5 days).              Analysis of AChE activity during the differentiation of SMDCs to multinucleated myotubes.           

Conclusion

The CytoSMART™ System in combination with Innovacell’s potency assay can provide detailed information on the dynamic process of SMDC differentiation as well as quantify myogenic potency of the tested cells. Furthermore, the correlation of cell-cell fusion and AChE activity within the cells becomes visible.  

 

References

  1. Frudinger A, Kolle D, Schwaiger W, Pfeifer J, Paede J, and Halligan S (2009) Gut 59, 01, 55–61.
  2. Frudinger A, Pfeifer J, Paede J, Kolovetsiou-Kreiner V, Marksteiner R, and Halligan S (2015) Colorectal Dis 17, 9, 794–801.
  3. Marksteiner R (2014) WO2014044867.
  4. Mauro A (1961) J Biophys Biochem Cytol 9, 493–495.
  5. Yin H, Price F, and Rudnicki M (2013) Physiol Rev 93, 1, 23–67.
  6. Ellman GL, Courtney KD, Andres JR V, and Feather-Stone RM (1961) Biochem Pharmacol 7, 88–95.

 

Find out more about Innovacell Biotechnology.

 

Interested to present your research here? Then apply to become a CytoSMART™ Lab. You will receive a CytoSMART™ System on loan for 4 weeks and your project will be featured on the Lonza website.

 

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