Our lab is differentiating iPSCs into cardiomyocytes and we have initial success for a few times then our efficiency drops. The only thing that is changing is cell density. Do you think the cell density is affecting our efficiency and if so, what kind of passaging do you recommend for the best efficiency?

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Accepted Answer

iPSC quality of the starter population and seeding densities are critical for cardiac differentiation depending on the protocol you are adopting for your differentiation to cardiomyocytes. If you have expanding populations in the beginning during differentiation, the stochastic ratios alter and could lead to less robust methods of generating desired cell types.  Similarly if the starting quality of iPSCs is poor, it will eventually result in poor differentiation of desired lineages.

One product you could try to improve your efficiency is the new PSC Cardiomyocyte Differentiation kit offered by Life Technologies. This kit offers a simple and reproducible three-step protocol to generate cardiomyocytes. Early cardiomyocytes can be generated as early as eight days and can be matured for the length of time required. Cells differentiated using this kit are applicable to studies including cardiotox screening, cardiac development, disease modeling, etc.

Learn more about the PSC Cardiomyocyte Differentiation Kit - http://www.lifetechnologies.com/us/en/home/life-science/stem-cell-research/stem-cell-differentiation/culture-systems-reagents-cardiomyocyte-differentiation.html

Answer

I am looking for a system that can detect differentiation using imaging techniques that are not invasive to the culture. Do you have any recommendations?

I am trying to culture human iPSCs without a feeder layer and keep them undifferentiated. I am using a commercial iPSCs media. I am trying to do this without matrigel, but the cells keep differentiating. Do I need to add Matrigel or is there a way to do this without?

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