In my lab we are using a published protocol for generating insulin producing beta cells. We are seeing significant variability, but am not sure if it is the protocol or some other factor. What is the level of variability you find with your kit and how simple would it be to train technicians to use this method instead? Is it easier than currently published protocols?

Answer

The STEMdiff™ Pancreatic Progenitor Kit is comprised of 2 basal media and 6 supplements that are used in specific combinations during the 2 week differentiation. Accompanying the kit is a detailed step-by-step protocol, including a schematic of the differentiation process. Because the medium is supplied in this modular format, there is no need for complex medium manufacturing on the part of the user. Simply mix the appropriate stage-specific supplement and basal medium and add it to your cells each day to achieve robust and efficient differentiation. With our own in house testing, we find that variability is quite low between experiments and across cell lines. For example, we measured the percent PDX-1⁺/NKX6.1⁺ cells at the end of Stage 4 across 10 different experiments using the H1 cell line. The data show an average differentiation efficiency of 69.3 ± 11.9% (mean ± SD) with a range of 46.9 – 83.3%. Within a given experiment, duplicate wells showed an average difference of 1.9 ± 1.2% (mean ± SD). Finally, we show that the average efficiency of differentiation is similar in human embryonic stem cell lines and induced pluripotent stem cell lines. For these data, please refer to Figure 2B here. For the kit protocol, please view this product information sheet.