Flow cytometry analysis of PrPc knockdown by siRNA in hDPSCs

Stem cells can develop into multiple cell types, from neurons to epithelial cells, through a process called “differentiation”. The  cellular form of prion protein (PrP^C), a cell surface glycoprotein located within lipid rafts, has been reported to be involved in various cellular pathways and could also have a role in the differentiation of stem cells into neurons.

Several studies have shown that the prion protein PrP^C may be subjected to post-translational cleavage events which produce active fragments such as N1, N2 or shed PrP^C. that seem to be implied in signalisation processes. Here, we examine the impact of shed  PrPC in signal pathways and in the neuronal differentiation process of stem cells using human dental pulp stem cells (hDPSCs) as a cellular model system. The recombinant prion protein 23–231 (recPrP^C ) is used to mimic the effects of shed PrP^C. The role of endogenous PrP^C in the neuronal differentiation process is evaluated using siRNA PrP to suppress the expression of prion protein PrP^C.

To verify whether the knockdown of PrP^C in hDPSCs has been effective, the cells were analyzed with flow cytometry using mouse anti-PrP SAF32 mAb (Bertin Technologies, Montigny-le-Bretonneux, France).