>>665 Jay Schufman said: Unfortunately, this sounds more like alchemy than science! If it is science, then it should be reproducible by following a set procedure, and that procedure should be elegant, not crouched in symantics and mysticism! (There’s one born every minute!)
Despite a media frenzy, especially in Japan, with headlines even suggesting the results are fraudulent, the authors have stood by the work. Today Niwa told Nature News that members of the team besides Obokata have replicated the bulk of the work and that others outside the laboratory have succeeded in the first critical step, inducing Oct3/4 expression after the acid treatment.
But the authors admit that the procedure is more complicated than originally advertised, leading to the publication of the tips.
The 10-page document states: “Despite its seeming simplicity, this procedure requires special care in cell handling and culture conditions, as well as in the choice of starting cell population.” The authors also point to the importance of bringing the cells gradually to the brink of death ― which kills some 80% of them after two to three days ― in order to reach the “optimal level of sub-lethal stress”.
The tips break down the process into three sections: collection of tissue and treatment with low-pH needed to produce STAP cells; preparing the culture needed to convert STAP cells to STAP stem cells which behave like iPS cells or embryonic stem cells; and preparing the culture needed to turn STAP cells into “FI cells”, which can form placenta.
The document includes 28 “important” tips, which note the necessity of starting with primary cells (as opposed to cultured cells) (1, ii); that mice less than a week old, especially male mice, gave better results (1, iv); the recommendation of using non-adhesive plates, which allow cell mobility and cluster formation (6, i); the importance of getting the cell density right in culture; the recommendation of using mice of a specific genetic lineage (8i), and many other detailed hints for using the proper culture conditions.