On 2014 Oct 16, Jongpil Kim commented:

To Whom It May Concern:

We have reviewed the concerns regarding some of the images presented in our study brought up by Paul Brookes on PubMed and Derek Lowe on his blog. In the wake of numerous controversial studies in the stem cell field, we appreciate the scrutiny of our results as skepticism plays a critical role in the progression of scientific research.

We have addressed all of the concerns raised on the internet regarding duplication of images and cropping of western blots and detail these responses separately.

First, regarding the similar radial contrast patterns in Figure 1C and 2D when black images are overexposed, we now provide high magnification versions of these images showing that while the radial contrast is similar between images (as would be expected when taking images from an essentially blank field on the same microscope with the same detector), there are clear differences observable upon higher magnification, demonstrating that these blank images were acquired independently.

Also, we have now provided all of the original western blots to the Journal that can be published in an erratum if the Journal deems this necessary. There were no sign of splicing or cropping of these original western data in Figure 4C, and we cut out the unnecessary part of images for the display on the limited area of paper in Figure 4F.

Finally, with regard to the image duplication of the brightfield micrograph of proliferating fibroblasts in Figure 2b: Yes, these are the same image, because this is the Time 0 timepoint of the experiment, prior to induction of EMF and factor infection. This began as a single culture at Time 0, after which half of the culture was exposed to EMF and the other half (control) was not (all in the presence of the Yamanaka factors). It is unclear to us why this is a concern for Paul Brookes, as he asserts this image was used to ‘represent different conditions’, however at Time 0, prior to the induction of EMF, these are in fact the same condition.

We are more than happy to provide all original images and discussion regarding these points in an erratum to the manuscript if the Journal deems it to be necessary.

On another note, we would like to point out significant errors made by Paul Brookes and Derek Lowe in their respective blogs in their interpretation of our study. Brookes states “in the wake of the STAP stem cell controversy, a paper claiming that iPSCs can be made using magnetic fields deserves special scrutiny”, and Lowe states “Yep, this paper says that stem cells can be produced from ordinary somatic cells by exposure to electromagnetic fields”. To us this suggests that neither Brookes nor Lowe actually read our manuscript, and rather just began altering contrast of images in the paper to find what they perceive as discrepancies.

Nowhere in our manuscript do we claim “iPSCs can be made using magnetic fields”. This would be highly suspect indeed. Rather, we demonstrate that in the context of highly reproducible and well-established reprogramming to pluripotency with the Yamanaka factors (Oct4, Sox2, Klf4, and cMyc/or Oct4 alone), EMF influences the efficiency of this process. Such a result is, to us, not surprising given that EMF has long been noted to have effects on biological system(Adey 1993, Del Vecchio et al. 2009, Juutilainen 2005)(There are a thousand of papers for biological effects of EMF on Pubmed) and given that numerous other environmental parameters are well-known to influence reprogramming by the Yamanaka factors, including Oxygen tension (Yoshida et al. 2009), the presence of Vitamin C (Esteban et al. 2010), among countless other examples.

For individuals such as Brookes and Lowe to immediately discount the validity of the findings without actually attempting to reproduce the central experimental finding is not only non-scientific, but borders on slanderous. We suggest that these individuals take their skepticism to the laboratory bench so that something productive can result from the time they invest prior to their criticizing the work of others.

“Often those that criticize others reveal what he himself lacks.” ― Shannon L. Alder

http://i.imgur.com/XgKBX47.jpg?1

http://i.imgur.com/gvZdZAQ.jpg

http://i.imgur.com/ljqPnxU.jpg

http://i.imgur.com/dEB1aoX.jpg

JP Kim

JONGPIL KIM, Ph.D. Assistant Professor Dept of Biomedical Engineering Dongguk University Seoul, Korea Tel: 82-02-2260-3371 Fax: 82-02-2260-8726 Email: jpkim153@dongguk.edu, jk2316@gmail.com

On 2014 Oct 14, Paul Brookes commented:

Cross-posted from PubPeer (https://pubpeer.com/publications/A730F960943331E553B54335FE8877)

This paper came up on Derek Lowe's "In the Pipeline" blog... http://pipeline.corante.com/archives/2014/10/14/electromagnetic_production_of_stem_cells_really.php

In the wake of the STAP stem cell controversy, a paper claiming that iPSCs can be made using magnetic fields deserves special scrutiny. Not surprisingly, there are a few problems with this paper...

Figure 2B - image re-use to represent different conditions http://i.imgur.com/Tu0kA52.jpg

Figures 4C & 4F - undisclosed splicing together of blots revealed at high contrast http://i.imgur.com/YREpQeF.jpg

Figures 1C & 4D - background fluo' images in some controls are "more similar than would be anticipated by pure coincidence" http://i.imgur.com/VKfgNIx.jpg http://i.imgur.com/2FfvvpH.jpg

There is another minor issue regarding the phase contrast images that are provided for numerous fluorescence images, including in the supplemental data. In short, it does not seem feasible that the fluorescent images shown could have originated from the cells shown in the phase contrast image. This probably means they just used different cells for the phase and the fluo' imaging, but this was not clearly stated anywhere, and is certainly not standard practice.

On 2014 Oct 14, Paul Brookes commented:

Cross-posted from PubPeer (https://pubpeer.com/publications/A730F960943331E553B54335FE8877)

This paper came up on Derek Lowe's "In the Pipeline" blog... http://pipeline.corante.com/archives/2014/10/14/electromagnetic_production_of_stem_cells_really.php

In the wake of the STAP stem cell controversy, a paper claiming that iPSCs can be made using magnetic fields deserves special scrutiny. Not surprisingly, there are a few problems with this paper...

Figure 2B - image re-use to represent different conditions http://i.imgur.com/Tu0kA52.jpg

Figures 4C & 4F - undisclosed splicing together of blots revealed at high contrast http://i.imgur.com/YREpQeF.jpg

Figures 1C & 4D - background fluo' images in some controls are "more similar than would be anticipated by pure coincidence" http://i.imgur.com/VKfgNIx.jpg http://i.imgur.com/2FfvvpH.jpg

There is another minor issue regarding the phase contrast images that are provided for numerous fluorescence images, including in the supplemental data. In short, it does not seem feasible that the fluorescent images shown could have originated from the cells shown in the phase contrast image. This probably means they just used different cells for the phase and the fluo' imaging, but this was not clearly stated anywhere, and is certainly not standard practice.

On 2014 Oct 16, Jongpil Kim commented:

To Whom It May Concern:

We have reviewed the concerns regarding some of the images presented in our study brought up by Paul Brookes on PubMed and Derek Lowe on his blog. In the wake of numerous controversial studies in the stem cell field, we appreciate the scrutiny of our results as skepticism plays a critical role in the progression of scientific research.

We have addressed all of the concerns raised on the internet regarding duplication of images and cropping of western blots and detail these responses separately.

First, regarding the similar radial contrast patterns in Figure 1C and 2D when black images are overexposed, we now provide high magnification versions of these images showing that while the radial contrast is similar between images (as would be expected when taking images from an essentially blank field on the same microscope with the same detector), there are clear differences observable upon higher magnification, demonstrating that these blank images were acquired independently.

Also, we have now provided all of the original western blots to the Journal that can be published in an erratum if the Journal deems this necessary. There were no sign of splicing or cropping of these original western data in Figure 4C, and we cut out the unnecessary part of images for the display on the limited area of paper in Figure 4F.

Finally, with regard to the image duplication of the brightfield micrograph of proliferating fibroblasts in Figure 2b: Yes, these are the same image, because this is the Time 0 timepoint of the experiment, prior to induction of EMF and factor infection. This began as a single culture at Time 0, after which half of the culture was exposed to EMF and the other half (control) was not (all in the presence of the Yamanaka factors). It is unclear to us why this is a concern for Paul Brookes, as he asserts this image was used to ‘represent different conditions’, however at Time 0, prior to the induction of EMF, these are in fact the same condition.

We are more than happy to provide all original images and discussion regarding these points in an erratum to the manuscript if the Journal deems it to be necessary.

On another note, we would like to point out significant errors made by Paul Brookes and Derek Lowe in their respective blogs in their interpretation of our study. Brookes states “in the wake of the STAP stem cell controversy, a paper claiming that iPSCs can be made using magnetic fields deserves special scrutiny”, and Lowe states “Yep, this paper says that stem cells can be produced from ordinary somatic cells by exposure to electromagnetic fields”. To us this suggests that neither Brookes nor Lowe actually read our manuscript, and rather just began altering contrast of images in the paper to find what they perceive as discrepancies.

Nowhere in our manuscript do we claim “iPSCs can be made using magnetic fields”. This would be highly suspect indeed. Rather, we demonstrate that in the context of highly reproducible and well-established reprogramming to pluripotency with the Yamanaka factors (Oct4, Sox2, Klf4, and cMyc/or Oct4 alone), EMF influences the efficiency of this process. Such a result is, to us, not surprising given that EMF has long been noted to have effects on biological system(Adey 1993, Del Vecchio et al. 2009, Juutilainen 2005)(There are a thousand of papers for biological effects of EMF on Pubmed) and given that numerous other environmental parameters are well-known to influence reprogramming by the Yamanaka factors, including Oxygen tension (Yoshida et al. 2009), the presence of Vitamin C (Esteban et al. 2010), among countless other examples.

For individuals such as Brookes and Lowe to immediately discount the validity of the findings without actually attempting to reproduce the central experimental finding is not only non-scientific, but borders on slanderous. We suggest that these individuals take their skepticism to the laboratory bench so that something productive can result from the time they invest prior to their criticizing the work of others.

“Often those that criticize others reveal what he himself lacks.” ― Shannon L. Alder

http://i.imgur.com/XgKBX47.jpg?1

http://i.imgur.com/gvZdZAQ.jpg

http://i.imgur.com/ljqPnxU.jpg

http://i.imgur.com/dEB1aoX.jpg

JP Kim

JONGPIL KIM, Ph.D. Assistant Professor Dept of Biomedical Engineering Dongguk University Seoul, Korea Tel: 82-02-2260-3371 Fax: 82-02-2260-8726 Email: jpkim153@dongguk.edu, jk2316@gmail.com