Quantification of COX-1 & COX-2 Expression from iPSC Derived Neuroprogenitor Cells

The applications of induced pluripotent stem cell (iPSC) research are reaching new depths as scientists make new discoveries about homeostasis and disease. Stem cell research has implications in many areas, from identification of disease mechanisms on a cellular level to development of treatments. A vital part of this research is focused on molecular identification of relevant genes. With that said, it is of utmost importance that researchers are able to obtain RNA from cultured iPSC’s that are not only high yield, but also not overly sheared in order to conduct downstream studies like RT-qPCR, RNA-seq, and other RNA-based analyses.

More specifically, serving as a drug target for pain attenuation and topic of study for involvement in various inflammatory disease mechanisms, cyclooxygenase enzymes (COX) are known to convert arachidonic acid to prostaglandin in the tissues, and central and peripheral nervous systems (Rouzer, 2009). Acting in response to many inflammatory pathways, both isoforms of the COX enzymes serve as sites for selective and non-selective inhibition for non-steroidal anti-inflammatory drugs as well as targets for study in the case of pathophysiologic anomalies.

Herein, we evaluate the efficacy of the Omni Bead Ruptor Elite™ bead mill homogenizer in front end sample preparation of cultured iPSC derived neural progenitor (NP) cells for isolation and purification of RNA, and detection of COX-1 and COX-2 via RT-qPCR.

Table 2: NP cell RNA concentration and Integrity Values.