We recently finished our Ask the Expert discussion on “Ask the Expert – Using Luminex Technology to Non-Invasively Monitor Stem Cell Differentiation”. During this Ask the Expert session, we discussed methods for characterizing stem cells and predicting differentiation. Specific topics included using the proteome profiler and Luminex multi-analyte assays, effects of media ingredients on secretion profiles, confirming pluripotency, and improving differentiation success.

One of the challenges facing Stem Cell Research and Cell Therapy applications today is the characterization and investigation of stem cell and differentiated populations. There are several options currently available for this type of investigation, for example, rt-PCR, western blot, immunochemistry, and flow cytometry. However all these methods are invasive, which means that valuable cells are lost during the process of characterization.

Recently, we published a Poster Article on “Predicting Differentiation and Characterizing Pluripotent Stem Cells Using Non-invasive Multi-analyte Luminex® Assays.” The article discusses a non-invasive method for cell characterization during differentiation. This method utilizes the flexibility of Proteome Profiler™ Antibody Arrays and Luminex® Assays to assess the expression of multiple analytes in cell culture supernates. Because this method uses cell culture supernates it is non-invasive and enables the detection of changes in cytokine expression throughout the progression of stem cell differentiation.

In this Ask the Expert session, Dr. Joy Aho, Manager of Product Development for the Stem Cell and Cell Culture Department at R&D Systems, answered your questions regarding using Luminex Technology to monitor stem cell differentiation without using any invasive methods, including using this technology to assess secretion profiles and how those differences may provide a method for predicting differentiation success and cell line specific optimization of differentiation protocols. Dr. Aho received a Ph.D. in Molecular, Cellular, Developmental Biology and Genetics from the University of Minnesota – Twin Cities and has over 10 years of experience developing media, reagents, and differentiation kits for stem cells.

Below is a sneak peek of the discussion, for a full transcript, please see – Ask the Expert – Ask the Expert – Using Luminex Technology to Non-Invasively Monitor Stem Cell Differentiation.

Question:

Can this methodology be applied to other stem cell types, like mesenchymal stem cells?

The Answer:

Most certainly. These techniques are applicable across a variety of different stem cells and differentiation protocols. All cells are secreting protein all of the time and it’s just a matter of using these methodologies to identify particular proteins of interest. From that point, for you can build secretory signatures across the differentiation time course of your stem cell of interest.

Question:

Using your method, can I confirm that my cells are still pluripotent without damaging them after reprogramming and during expansion?

The Answer:

In order to non-invasively monitor pluripotency, you would first need to determine the signature secretory profile of your pluripotent stem cells. I’d suggest first assessing the secretory profile of your cells with the qualitative Proteome Profile XL Cytokine Array. You can compare your results with the data presented in the webinar, for which we show similar secretory profiles across iPS cell lines. As an alternate, there are pluripotent live-cell staining kits available that allow you to visually confirm pluripotency in live cells using fluorescence without out affecting proliferation or stemness.

Question:

Do you have any customers who have been able to use this method to improve their differentiation success rate?

The Answer:

The use of non-invasive techniques, such as Luminex and Proteome Profiler Arrays, to monitor stem cell differentiation is relatively new. Our initial studies have focused on profiling differentiation of iPSCs into various lineages. We are currently working in-house and with collaborators on ways in which this technology could identify growth factors or cytokines that could be added to improve differentiation success. We are interested in more collaborative opportunities to explore the boundaries of these platforms for predicting and optimizing stem cell differentiation.

For more information on this technology, please view Dr. Aho’s webinar “How to Non-invasively Monitor Stem Cell Differentiation using Luminex Technology,” now available on demand.

As Stem Cell Research moves closer toward therapeutic application, there is a recognized need to increase the quality and consistency of pluripotent stem cells and their derivatives. While variability in the efficiency of stem cell differentiation can undermine data interpretation and slow research progress, the ability to quickly monitor changes in cell phenotype without sacrificing yield continues to be a challenge. In this webinar, Richard Feurstenberg and Joy Aho discuss Luminex technology, its power as a non-invasive tool for monitoring stem cell differentiation, and its utility for troubleshooting and optimizing stem cell culturing and differentiation conditions.