We have found this to be an effective protocol for generating STAP cells in our lab, regardless of the cell type being studied. The protocol below is a combination of the two most effective approaches described in our Jan 31, 2014 article published in Nature (Obokata et al., Stimulus Triggered Fate Conversion of Somatic Cells into Pluripotency. Nature 505. 641-647, 2014). It is very important that each step be performed precisely as described. The protocol is extremely simple, but will vary slightly if you are starting with tissue rather than a cell suspension. It also will vary depending upon the cell type or tissue with which you are starting. It is important to not skip any steps. It is especially important to triturate the cell suspension for a minimum of 30 minutes, until the suspension can be easily triturated up and down the reduced bore pipettes of the smallest orifices. We first describe the protocol when starting with a suspension of cells, and then describe additional steps necessary when starting with a soft tissue.

PROTOCOL FOR GENERATING STAP CELLS FROM MATURE SOMATIC CELLS.pdf