preprint published Momoko Watanabe author Jillian R. Haney author Neda Vishlaghi author Felix Turcios author Jessie E. Buth author Wen Gu author Amanda J. Collier author Osvaldo Miranda author 862A3C56-A8BF-11E9-B4FA-D9E3E56974250000-0001-6618-6889 Di Chen author Shan Sabri author Amander T. Clark author Kathrin Plath author Heather R. Christofk author Michael J. Gandal author Bennett G. Novitch author Telencephalic organoids generated from human pluripotent stem cells (hPSCs) are emerging as an effective system to study the distinct features of the developing human brain and the underlying causes of many neurological disorders. While progress in organoid technology has been steadily advancing, many challenges remain including rampant batch-to-batch and cell line-to-cell line variability and irreproducibility. Here, we demonstrate that a major contributor to successful cortical organoid production is the manner in which hPSCs are maintained prior to differentiation. Optimal results were achieved using fibroblast-feeder-supported hPSCs compared to feeder-independent cells, related to differences in their transcriptomic states. Feeder-supported hPSCs display elevated activation of diverse TGFβ superfamily signaling pathways and increased expression of genes associated with naïve pluripotency. We further identify combinations of TGFβ-related growth factors that are necessary and together sufficient to impart broad telencephalic organoid competency to feeder-free hPSCs and enable reproducible formation of brain structures suitable for disease modeling. Cold Spring Harbor Laboratory2019 eng 10.1101/2019.12.13.875773 75 yes Watanabe M, Haney JR, Vishlaghi N, et al. TGFβ superfamily signaling regulates the state of human stem cell pluripotency and competency to create telencephalic organoids. <i>bioRxiv</i>. 2019. doi:<a href="https://doi.org/10.1101/2019.12.13.875773">10.1101/2019.12.13.875773</a> Watanabe M, Haney JR, Vishlaghi N, Turcios F, Buth JE, Gu W, Collier AJ, Miranda O, Chen D, Sabri S, Clark AT, Plath K, Christofk HR, Gandal MJ, Novitch BG. 2019. TGFβ superfamily signaling regulates the state of human stem cell pluripotency and competency to create telencephalic organoids. bioRxiv, <a href="https://doi.org/10.1101/2019.12.13.875773">10.1101/2019.12.13.875773</a>. Watanabe, Momoko, Jillian R. Haney, Neda Vishlaghi, Felix Turcios, Jessie E. Buth, Wen Gu, Amanda J. Collier, et al. “TGFβ Superfamily Signaling Regulates the State of Human Stem Cell Pluripotency and Competency to Create Telencephalic Organoids.” <i>BioRxiv</i>. Cold Spring Harbor Laboratory, 2019. <a href="https://doi.org/10.1101/2019.12.13.875773">https://doi.org/10.1101/2019.12.13.875773</a>. M. Watanabe, J.R. Haney, N. Vishlaghi, F. Turcios, J.E. Buth, W. Gu, A.J. Collier, O. Miranda, D. Chen, S. Sabri, A.T. Clark, K. Plath, H.R. Christofk, M.J. Gandal, B.G. Novitch, BioRxiv (2019). M. Watanabe <i>et al.</i>, “TGFβ superfamily signaling regulates the state of human stem cell pluripotency and competency to create telencephalic organoids,” <i>bioRxiv</i>. Cold Spring Harbor Laboratory, 2019. Watanabe, Momoko, et al. “TGFβ Superfamily Signaling Regulates the State of Human Stem Cell Pluripotency and Competency to Create Telencephalic Organoids.” <i>BioRxiv</i>, Cold Spring Harbor Laboratory, 2019, doi:<a href="https://doi.org/10.1101/2019.12.13.875773">10.1101/2019.12.13.875773</a>. Watanabe, M., Haney, J. R., Vishlaghi, N., Turcios, F., Buth, J. E., Gu, W., … Novitch, B. G. (2019). TGFβ superfamily signaling regulates the state of human stem cell pluripotency and competency to create telencephalic organoids. <i>bioRxiv</i>. Cold Spring Harbor Laboratory. <a href="https://doi.org/10.1101/2019.12.13.875773">https://doi.org/10.1101/2019.12.13.875773</a> 73582020-01-23T09:53:40Z2022-06-17T08:03:32Z