Reply to: “Correspondence to bipolar disorder-iPSC derived neural progenitor cells exhibit dysregulation of store-operated Ca2+ entry and accelerated differentiation” by Yde Ohki and colleagues

Abstract

We appreciate the opportunity to respond to Yde Ohki and colleagues’ commentary [1] on our recent publication [2]. We agree with the authors that sufficient and accurate information is critical for experimental reproducibility. Therefore, we added the following missing details and corrected typographical errors in our paper. First, regarding the YFP-STIM1/2 puncta formation assay that we present in Fig. 2I–K, it should have been mentioned in the Puncta Formation Assay section of the Supplementary Information that the pEX-CMV-SP-YFP-STIM1 (Plasmid #18857, initially described in ref. [3]) and pEX-CMV-SP-YFP-STIM2 (Plasmid #18862, initially described in ref. [4]) constructs were purchased from Addgene. Furthermore, 2.5 µg plasmid DNA and 12.5 µL of Lipofectamine 2000 reagent were used per dish for each transfection. Second, in the Neurosphere Assay section of the Supplementary Information, we incorrectly wrote that the …[iPSC/NPCs] were agitated at 75 rpm in NEM for 72 h. This should have been 48 h, as illustrated in Fig. 5A of our paper. And third, two typos concerning cell line ID were identified by Ohki and colleagues: in the legend and caption of Supplementary Fig. S8 we mistakenly wrote PSC-01-121 when this line was PSC-01-122, and we incorrectly wrote GM05440 when it should have been GM05224 in the NanoString miRNA Profiling method section and last row of the Reagent or Resource table in the Supplementary Information. One mention of GM05440 was also corrected to GM05224 in the main paper. Those corrections have been made to the available online documents. We would also like to highlight that p-values were always corrected in the case of multiple comparisons, such as the RNA-sequencing dataset. That information can be found in the Supplementary Methods. We thank the authors for bringing these oversights to our attention and regret any confusion they may have caused.

The six cell lines (3 bipolar disorder patients and 3 healthy controls) that we characterized in our study (PSC-01-024, PSC-01-223, PSC-01-185, PSC-01-003, PSC-01-009, PSC-01-122) were generated as part of NIMH Project Number 5P50MH106933-02 (NIH RePORTER, Neuropsychiatric Genome-Scale and RDOC Individualized Domains, https://reporter.nih.gov/project-details/8929310) and are available through the NIMH Repository & Genomics Resource (Study 163, https://studyreg.nimhgenetics.org/ListOfStudies.jsp). Those cell lines were used in a previous study [5]. Further, two additional cell lines from the Coriell Cell Repository (one healthy control GM8330 and one bipolar disorder GM05224) were included in our NanoString miRNA Profiling experiment. These two lines have been extensively published (for examples, see refs. [6,7,8,9,10,11]). Finally, the use of neural induction supplement to produce neural progenitor cells (NPCs) from human pluripotent stem cells has been used by many other studies (for examples, see refs. [12,13,14,15,16]). This approach, which does not require the intermediary step of embryoid body formation, helps scale up production and maintain cells at the NPC stage of development. This information has been added to the Supplementary Material and Methods online.