Thomson:  "For transplantation therapies based on these cells, with the exception of autoimmune diseases, patient-specific iPS cell lines should largely eliminate the concern of immune rejection. It is important to understand, however, that before the cells can be used in the clinic, additional work is required to avoid vectors that integrate into the genome, potentially introducing mutations at the insertion site. ... However, further work is needed to determine if human iPS cells differ in clinically significant ways from ES cells" (emphases added).

As has already been suggested above, if there are so many "unknown" genetic mutations, antigens, or artifacts, there are not only obvious concerns about the use of these "patient-specific" cells in patients, but also about any validity of the data derived from "studies of disease mechanisms" and from screening drugs, vaccines, chemical and biological agents, etc.

V. Other possible sources of artifacts and foreign antigens

Although not specifically addressed in these studies, there are a host of other possible sources of artifacts and foreign antigens that could render the data presented as useless, as well as call into serious question the use of iPS cells in sick patients as "therapies". Although some of the following examples may be too "scientific" for many readers, at least it is worth registering and considering some of them.

In general, there are too many known and unknown variables throughout the experiments; in fact, there is, as noted in the studies, genetic variability even within the same iPS clone batches. It is also critically relevant to determine from the studies if the assay machines, gels, tests, etc., were reliably and properly calibrated before use, so that the data would be reliable and reproducible. Otherwise, the data presented in these studies are essentially useless. Again, the data derived from just one or two studies could hardly pass a t-test. It is not clear if the data presented in these studies were determined to be statistically significant or not.

Other possible sources of data artifacts and problematic antigens in these studies involves the viral genes used as vectors, as well as the transmission genes transferred. For example, Thomson used the following transmission genes, all of which would definitely show as problematic antigens on the surfaces of iPS cells produced with them: OCT4, SOX2, NANOG, and LIN28; Yamanaka used the following transmission genes: Oct3/4, Sox2, Klf4, and c-Myc. And the questions arises as to whether all of the transmission genes used in the studies were derived from genetically different human embryos or fetuses? That too could result in artifacts in the data presented, as well as the presence of more foreign antigens on the surfaces of the iPS cells produced.

Also to be considered are foreign genes or antigenic molecules that could be inadvertently derived from the media, inoculations and feeder cell layers used, as well as from impurities in the several manufactured biological products, laboratory contamination, etc.

The above are simply lists of standard good quality research controls used throughout the various research fields. It would be important, then, to turn again to the specific research studies to determine any specific sources of artifacts and antigens.

A. Yamanaka et al

As noted, Yamanaka et al used the following transmission genes: Oct3/4, Sox2, Klf4, and c-Myc. All of these genes, if incorporated into the iPS cells, would cause foreign genetic antigens on the iPS cell surfaces. As already noted, one also wonders if all of these "normal" transmission genes were derived from different human embryos or fetuses. One cannot assume, e.g., that the Oct3/4 transmission gene from one human embryo is genetically identical to that of other human embryos.  If such genetic differences exist, then they could cause additional foreign antigens on the surface of iPS cells.

Viral genes from the vectors used could also be incorporated into the genome of iPS cells, also causing foreign antigens on the cell surfaces.  For Yamanaka, all four retroviruses used to transmit the "normal" human embryonic genes could cause foreign antigens, not to mention specifically the amphotropic retrovirus, the lentivirus, and the ecotropic retrovirus used.  Viruses have genes, too!

And for brevity, the following is a further listing of sources of data artifacts, foreign antigens in the iPS cells produced, and human embryonic/fetal cell sources (at least as much as can be determined solely from the studies):

• the mouse receptor for retroviruses, Slc7a1, was injected into the original HDF cells
• firefly luciferase gene was injected as well
• one target cell used, HDF, was derived from the facial dermis of a 36-year-old Caucasian female, each detail of which would produce specific antigens on the iPS cell surface
• another target cell used was a primary human fibroblast-like synoviocytes (HFLS) from the synovial tissue of a 69-year-old Caucasian male, each detail of which would produce specific antigens on the iPS cell surface
• another cell used, BJ cells, has its own specific cell antigens, as well as being a cell line established from neonate fibroblasts
• green fluorescent protein (GFP) used
• mitomycin C-treated SNL feeder cells used
• medium DMEM containing 10% FBS (fetal source)
• medium for primate ES cell culture used
• medium for primate ES cell culture containing bFGF used
• fibroblast growth factor (bFGF) used
• MEF-conditioned primate ES cell medium used
• PA6 feeder layer used
• coculture with PA6 cells
• activin A and bone morphogenetic protein (BMP) used
• 0.5% penicillin and streptomycin in media used
• DMEM containing 10% FBS (fetal source) used
• L-glutamine (Invitrogen), 1 3 10_4 M nonessential amino acids (Invitrogen), 1 mM sodium pyruvate (Sigma) used
• PA6 stroma cells used
• Primate ES medium used
• recombinant human basic fibroblast growth factor (bFGF) used
• DMEM/F12 containing 1 mg/ml collagenase IV used
• 0.3 mg/ml Matrigel (growth-factor reduced) used
• MEFs derived from embryonic day 13.5 embryo pool of ICR mice used
• EcoRI fragment of pCR2.1-hOCT3/4 introduced into the EcoRI site of pMXs retroviral vector used
• "20 bp random sequence, designated N20 barcode, into the NotI/SalI site of Oct3/4 expression vector used; unique barcode sequence in each experiment"
• KpnI/BglII digestion
• AatII (blunted)/NheI fragment of pQBI-polII inserted into the KpnI (blunted)/NheI site of pGV-BM2
• Lentivirus production: 293FT cells (fetal cells), transfected with 3 mg of pLenti6/UbC-Slc7a1 along with 9 mg of Virapower packaging mix
• PLAT-E packaging cells; transfected with pMXs vectors with Fugene 6 transfection reagent
• 20% knockout serum replacement used
• PA6-feeder layer in Glasgow minimum essential medium used
• RPMI1640 (Invitrogen) plus B27 supplement (Invitrogen) medium (RPMI/B27), supplemented with 100 mg/ml human recombinant activin A used
• human recombinant bone morphologenic protein 4 used

B. Thomson

Whereas Yamanak's study provided many (but not all) details of their materials and methods, the same is not true for Thomson's study.  In fact, there was a dirth of "methods and procedures" details. Therefore, it is not possible to identify the same kind of list as above. However, it is still obvious that foreign genetic cell surface antigens would be caused in their experimental iPS cells because the cells were infected and transformed with foreign transmission genes (OCT4, SOX2, NANOG, and LIN28), using four different viral vectors whose own genes would be foreign.

VI. Conclusion

Considering the enormous stakes involved, it would seem that it is incumbent upon scientists involved in especially ethically sensitive research to be as open and detailed in their publications as possible. The trend, however, seems to be to camouflage and dilute the scientific details as much as possible -- not only in order to evade professionally appropriate questions from their scientific peers, but also to evade the very questions that society and ethics have traditionally required of all scientists. Rather than use and report the accurate scientific facts, it would seem that scientists prefer to secure their successes by using false and misleading manufactured "scientific" terms, verbal hype, and empty promises.

Even the most basic requirements of research ethics appear to have been abandoned, including the required use of the most accurate and reliable scientific facts, as well as the "denial" of the age-old dictum that it is simply wrong to purposefully kill innocent human beings - regardless of their stage of development, ethnicity, culture, degree of illness, etc., and regardless of whether or not their destruction could be of benefit to other human beings.

Needless to say, even the most desperate of sick patients should not be exposed to "therapeutic" research or clinical trials when such participation unwittingly puts them into serious danger of harm and even death. One even wonders how such patients could give legally valid "informed consent".

We would all obviously welcome a viable scientific and ethical resolution to the divisive politics of human embryo and fetal research that has consumed us for so many years. These iPS studies, however, do not appear to be that solution.