Stem Cell Core

Services Provided

The Stem Cell Core specializes in the generation, maintenance and differentiation of human iPS cells. The core has expertise in human iPS cell reprogramming and CRISPR-Cas9 genome editing of human iPS cells. The core supports researchers by providing the physical space, validated reagents, iPS cell lines and hands-on training for scientists to use iPS cells for their research goals.

Human iPS Cells Reprogramming

• iPS Cell Reprogramming from Human Peripheral Blood Mononuclear Cells (PBMCs) (Episomal method)
• iPS Cell Reprogramming from Fibroblasts (ReproRNA StemCell Technologies Kit)

Cardiomyocyte Differentiation from Human iPS Cells

• Modulating Wnt Signaling
• StemCell Technologies Kit

Cerebral Organoid Differentiation from Human iPS Cells

• StemCell Technologies Kit

Transcription Factor Mediated Differentiation of Human iPS Cells

• Cortical Neurons (by Inducing NGN2)
• Motor Neurons (by Inducing ISL1, LHX3 and NGN2)

Genome Engineering of Human iPS Cells

• CRISPR-Cas9 Gene Knockout by NHEJ
• CRISPR-Cas9 Gene Knock-in by HDR

Training Services

• Human iPS Cells Culture and Maintenance
• Cardiomyocyte Differentiation from Human iPS Cells
• Human iPS Cell Reprogramming from Fibroblasts and PBMCs

Supply of Cell Lines

• WTC-iPSC
• WTB-iPSC
• CRISPRi-iPSC
• CRISPRn-iPSC
• GCaMP6f-iPCS

Instrumentation

Fees and Scheduling

• Use of the Stem Cell Core facility is subject to fees charged as an add-on cost to in-house media purchased; alternatively, fees are charged via the purchase of a “sticker”. The add-on cost covers the use of plastics, PPE, some commonly used reagents, and standard equipment usage.
• Costs/fees for media, reagents, scientific services, and specialized equipment usage are listed on iLab. An iLab account is required to view specific pricing.
• The first 30-minute consultation is free of charge.
• For all fee inquiries or to set up an iLab account, email stemcell-core@gladstone.ucsf.edu.
• For custom scientific project inquiries or inquiries about use of specialized instruments, email stemcell-core@gladstone.ucsf.edu.

Scheduling

• To schedule new consultations and service requests, contact stemcell-core@gladstone.ucsf.edu. Once accepted, log into your Stem Cell Core iLab account and request the service.
• To schedule equipment use, visit iLab and book in the calendar.

Publications

Transcription Factor Protein Interactomes Reveal Genetic Determinants in Heart Disease. Barbara Gonzalez-Teran , Maureen Pittman , Franco Felix , Reuben Thomas , Desmond Richmond-Buccola , Ruth Hüttenhain , Krishna Choudhary , Elisabetta Moroni , Mauro W Costa , Yu Huang , Arun Padmanabhan , Michael Alexanian , Clara Youngna Lee , Bonnie E J Maven , Kaitlen Samse-Knapp , Sarah U Morton , Michael McGregor , Casey A Gifford , J G Seidman , Christine E Seidman , Bruce D Gelb , Giorgio Colombo , Bruce R Conklin , Brian L Black , Benoit G Bruneau , Nevan J Krogan , Katherine S Pollard , Deepak Srivastava Cell 2022 Mar 3;185(5):794-814.

Brahma Safeguards Canalization of Cardiac Mesoderm Differentiation. Swetansu K Hota , Kavitha S Rao , Andrew P Blair , Ali Khalilimeybodi , Kevin M Hu , Reuben Thomas , Kevin So , Vasumathi Kameswaran , Jiewei Xu , Benjamin J Polacco , Ravi V Desai , Nilanjana Chatterjee , Austin Hsu , Jonathon M Muncie , Aaron M Blotnick , Sarah A B Winchester , Leor S Weinberger , Ruth Hüttenhain , Irfan S Kathiriya , Nevan J Krogan , Jeffrey J Saucerman , Benoit G Bruneau. Nature 2022 Feb;602(7895):129-134.

Transcription Factor GATA4 Regulates Cell Type-Specific Splicing Through Direct Interaction With RNA in Human Induced Pluripotent Stem Cell-Derived Cardiac Progenitors. Lili Zhu , Krishna Choudhary , Barbara Gonzalez-Teran , Yen-Sin Ang , Reuben Thomas Nicole R Stone , Lei Liu , Ping Zhou , Chenchen Zhu , Hongmei Ruan , Yu Huang , Shibo Jin, Angelo Pelonero , Frances Koback , Arun Padmanabhan , Nandhini Sadagopan , Austin Hsu , Mauro W Costa , Casey A Gifford , Joke G van Bemmel , Ruth Hüttenhain , Vasanth Vedantham , Bruce R Conklin , Brian L Black , Benoit G Bruneau , Lars Steinmetz , Nevan J Krogan , Katherine S Pollard , Deepak Srivastava. Circulation 2022 Sep 6;146(10):770-787.

Sars-Cov-2 Infection Of Human Ipsc-Derived Cardiac Cells Reflects Cytopathic Features in Hearts of Patients with Covid-19. Perez-Bermejo JA, Kang S, Rockwood SJ, Simoneau CR, Joy DA, Silva AC, Ramadoss GN, Flanigan WR, Fozouni P, Li H, Chen PY, Nakamura K, Whitman JD, Hanson PJ, McManus BM, Ott M, Conklin BR, McDevitt TC. Sci Transl Med 2021 Apr 21;13(590).

Allele-Specific Gene Editing Rescues Pathology in a Human Model of Charcot-Marie-Tooth Disease Type 2E. Feliciano CM, Wu K, Watry HL, Marley CBE, Ramadoss GN, Ghanim HY, Liu AZ, Zholudeva LV, McDevitt TC, Saporta MA, Conklin BR, Judge LM. Front Cell Dev Biol 2021 Aug 16;9:723023.

Transcription Factor Overexpression Drives Reliable Differentiation of Retinal Pigment Epithelium From Human Induced Pluripotent Stem Cells. Dewell TE, Gjoni K, Liu AZ, Libby ARG, Moore AT, So PL, Conklin BR. Stem Cell Res 2021 May;53:102368.

Sox2 and Klf4 as the Functional Core in Pluripotency Induction without Exogenous Oct4. An, Z., Liu, P., Zheng, J., et al. 2019. Cell Reports 29(7), p. 1986–2000.e8.

Differentiation of V2a interneurons from human pluripotent stem cells. Butts, J.C., McCreedy, D.A., Martinez-Vargas, J.A., et al. 2017. Proceedings of the National Academy of Sciences of the United States of America 114(19), pp. 4969–4974.

Oligogenic inheritance of a human heart disease involving a genetic modifier. Gifford, C.A., Ranade, S.S., Samarakoon, R., et al. 2019. Science 364(6443), pp. 865–870.

BMP-SMAD-ID promotes reprogramming to pluripotency by inhibiting p16/INK4A-dependent senescence. Hayashi, Y., Hsiao, E.C., Sami, S., et al. 2016. Proceedings of the National Academy of Sciences of the United States of America 113(46), pp. 13057–13062.

Miniaturized iPS-Cell-Derived Cardiac Muscles for Physiologically Relevant Drug Response Analyses. Huebsch, N., Loskill, P., Deveshwar, N., et al. 2016. Scientific Reports 6, p. 24726.

Automated Video-Based Analysis of Contractility and Calcium Flux in Human-Induced Pluripotent Stem Cell-Derived Cardiomyocytes Cultured over Different Spatial Scales. Huebsch, N., Loskill, P., Mandegar, M.A., et al. 2015. Tissue Engineering. Part C, Methods 21(5), pp. 467–479.

A BAG3 chaperone complex maintains cardiomyocyte function during proteotoxic stress. Judge, L.M., Perez-Bermejo, J.A., Truong, A., et al. 2017. Journal of Clinical Investigation Insight 2(14). PMC5518554

Spatiotemporal mosaic self-patterning of pluripotent stem cells using CRISPR interference. Libby, A.R., Joy, D.A., So, P.-L., et al. 2018. eLife 7.

CRISPR Interference Efficiently Induces Specific and Reversible Gene Silencing in Human iPSCs. Mandegar, M.A., Huebsch, N., Frolov, E.B., et al. 2016. Cell Stem Cell 18(4), pp. 541–553.

Isolation of single-base genome-edited human iPS cells without antibiotic selection. Miyaoka, Y., Chan, A.H., Judge, L.M., et al. 2014. Nature Methods 11(3), pp. 291–293.

Unbiased detection of CRISPR off-targets in vivo using DISCOVER-Seq. Wienert, B., Wyman, S.K., Richardson, C.D., et al. 2019. Science 364(6437), pp. 286–289.

The Cellular NMD Pathway Restricts Zika Virus Infection and Is Targeted by the Viral Capsid Protein. Fontaine, K.A., Leon, K.E., Khalid, M.M, Tomar, S., Jimenez-Morales, D., Dunlap, M., Kaye, J.A., Shah, P.S., Finkbeiner, S., Krogan, N.J., Ott. M. 2018. mBio. Nov-Dec; 9(6) PMCID: PMC6222128

Efficient CRISPR/Cas9-Based Genome Engineering in Human Pluripotent Stem Cells. Kime, C., Mandegar, M. A., Srivastava, D., Yamanaka, S., Conklin, B. R., & Rand, T. A. (2016). Current Protocols in Human Genetics, 88, 21.4.1–21.4.23.

Human disease modeling reveals integrated transcriptional and epigenetic mechanisms of NOTCH1 haploinsufficiency. Theodoris, C. V., Li, M., White, M. P., Liu, L., He, D., Pollard, K. S., Bruneau, B. G., & Srivastava, D. (2015). Cell, 160(6), 1072–1086.

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