This an interesting project. As you know many groups have relied upon cell surface markers such as CD44 and CD24 to identify the CSC population. In this project you have employed a marker-free approach to isolate CSCs which is unique. What are the benefits of this method compared to Marker dependent isolation?
Excellent question, I think we first need to clarify why we tried to avoid sorting. We have a really high rate of apoptotic cells in any sort we tried from primary cancer tissue. You have to keep in mind that the cells already are in a mechanical/enzymatic digest for hours. Labelling and sorting just adds to the pile of dead cells. We wanted to circumvent that problem. So our approach relies on a "CSC-only environment" and it works out for us. Our conditions allow only CSCs to continuously proliferate and self-renew. Our cultures are relatively pure in CSC content (paper in the submission process). We can grow the cells for 30+ passages and they stay stable for that time. Of course we can also show recapitulation of primary tumor in limiting transplants. Our next publication will highlight the cells and method. Stay tuned ;-)
Dear Dr. Maurer thanks for your response. That's interesting. In other words, in 3D hypoxic model you try to mimic the true tumor microenvironment and in vivo condition. i am eagerly waiting for your upcoming article about this project.;)
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