Developmental Potency and Metabolic Traits of Extended Pluripotency Are Faithfully Transferred to Somatic Cells via Cell Fusion-Induced Reprogramming
, Yean-Ju Hong 1, Hyeonwoo La 1, Tae-Kyung Hong 1,2, Kwonho Hong 1 and Jeong-Tae Do 1,2,*
Round 1
Reviewer 1 Report
In the manuscript “Developmental potency and metabolic traits of extended pluripotency are faithfully transferred to somatic cells via cell fusion-induced reprogramming” Jae Hoon Song et al. demonstrated that murine neural stem cells (NSCs) can be reprogrammed to and extended pluripotent state by fusion with murine extended pluripotent stem cells (EPSCs). They showed that tetraploid hybrid cells exhibit similar EPSCs transcriptional profile, acquisition of extraembryonic developmental potential and mitochondrial morphological and metabolic remodeling to the EPSC state.
The manuscript is concise and well written. The experiments are robust and performed with all appropriate controls. Results are quite relevant as they demonstrate for the first time that EPSCs can transfer extended pluripotency to committed progenitors like NSCs. However, it remains as an open question if extended pluripotency can be transferred to terminal differentiated somatic cells like fibroblasts. Nevertheless, I understand that this issue exceeds the aims of this work.
Minor issues:
1- On Materials and Methods section, most superscripts and subscripts are missing. Example: 1 x 105 ESCs should be corrected to 1 x 105 ESCs; CO2 incubator should be changed to CO2 incubator.
2- Authors observed that expression levels of EPSC-specific genes were higher in the hybrid cells than in EPSCs (Figure 2F). It would be interesting if you they speculate in the discussion why they think this happens. Could be a consequence of the tetraploid status?
Author Response
Please see the attachment.
Author Response File: 
Author Response.docx
Reviewer 2 Report
In this manuscript, Jae Hoon Song and colleagues use EPSCs fused OG2+/- ROSA+/-neural stem cells to reprogram the somatic cells and found that fusion hybrid cells expressed pluripotential genes and contributed to the extraembryonic and embryonic lineages in vivo and in vitro. They also performed RNA-sequencing analysis and metabolic analysis to confirm that the hybrid cells showed distinct global expression patterns and metabolic profiles resembling EPSCs. The author demonstrated that the extended pluripotency of EPSCs could be transferred to somatic cells through cell fusion-induced reprogramming. This research is very interesting. The manuscript is well written in English. The method is in detail and the results are clear. Here are my minor points.
1. Please describe the origin of OG2+/-/ROSA26+/- double transgenic NSCs
2. Figure 2D: No scale bar
3. Figure 2E: Nanog and DAPI didn’t match.
4. Figure 2F: The asterisk should be labeled on the top of the hybrid cells group
5. Figure 4: Gene names of the heatmap are not clear.
6. Figure 6 F: Gene names of the heatmap are not clear.
Author Response
Please see the attachment.
Author Response File: Author Response.docx
