iScience (Apr 2022)
A dual SHOX2:GFP; MYH6:mCherry knockin hESC reporter line for derivation of human SAN-like cells
- Zaniar Ghazizadeh,
- Jiajun Zhu,
- Faranak Fattahi,
- Alice Tang,
- Xiaolu Sun,
- Sadaf Amin,
- Su-Yi Tsai,
- Mona Khalaj,
- Ting Zhou,
- Ryan M. Samuel,
- Tuo Zhang,
- Francis A. Ortega,
- Miriam Gordillo,
- Dorota Moroziewicz,
- Daniel Paull,
- Scott A. Noggle,
- Jenny Zhaoying Xiang,
- Lorenz Studer,
- David J. Christini,
- Geoffrey S. Pitt,
- Todd Evans,
- Shuibing Chen
Affiliations
- Zaniar Ghazizadeh
- Department of Surgery, Weill Cornell Medical College, New York, NY 10065, USA; Corresponding author
- Jiajun Zhu
- Department of Surgery, Weill Cornell Medical College, New York, NY 10065, USA
- Faranak Fattahi
- The Center for Stem Cell Biology, Memorial Sloan Kettering Cancer Center, New York, NY 10065, USA; Department of Cellular and Molecular Pharmacology, University of California, San Francisco, San Francisco, CA 94158, USA; Eli and Edythe Broad Center of Regeneration Medicine and Stem Cell Research, University of California, San Francisco, San Francisco, CA 94143, USA
- Alice Tang
- Department of Surgery, Weill Cornell Medical College, New York, NY 10065, USA
- Xiaolu Sun
- Cardiovascular Research Institute, Weill Cornell Medicine, New York, NY 10021, USA
- Sadaf Amin
- Department of Surgery, Weill Cornell Medical College, New York, NY 10065, USA
- Su-Yi Tsai
- Department of Life Science, National Taiwan University, Taipei 10617, Taiwan
- Mona Khalaj
- Department of Surgery, Weill Cornell Medical College, New York, NY 10065, USA
- Ting Zhou
- The Center for Stem Cell Biology, Memorial Sloan Kettering Cancer Center, New York, NY 10065, USA
- Ryan M. Samuel
- Department of Cellular and Molecular Pharmacology, University of California, San Francisco, San Francisco, CA 94158, USA; Eli and Edythe Broad Center of Regeneration Medicine and Stem Cell Research, University of California, San Francisco, San Francisco, CA 94143, USA
- Tuo Zhang
- Genomic Resource Core Facility, Weill Cornell Medical College, New York, NY 10065, USA
- Francis A. Ortega
- Physiology, Biophysics, and Systems Biology Graduate Program, Weill Cornell Medical College, New York, NY 10065, USA; Department of Physiology & Pharmacology, SUNY Downstate Health Sciences University, Brooklyn, NY 11203, USA
- Miriam Gordillo
- Department of Surgery, Weill Cornell Medical College, New York, NY 10065, USA
- Dorota Moroziewicz
- The New York Stem Cell Foundation Research Institute, 619 West 54th Street, 3rd Floor, New York, NY 10019, USA
- Daniel Paull
- The New York Stem Cell Foundation Research Institute, 619 West 54th Street, 3rd Floor, New York, NY 10019, USA
- Scott A. Noggle
- The New York Stem Cell Foundation Research Institute, 619 West 54th Street, 3rd Floor, New York, NY 10019, USA
- Jenny Zhaoying Xiang
- Genomic Resource Core Facility, Weill Cornell Medical College, New York, NY 10065, USA
- Lorenz Studer
- The Center for Stem Cell Biology, Memorial Sloan Kettering Cancer Center, New York, NY 10065, USA
- David J. Christini
- Department of Physiology & Pharmacology, SUNY Downstate Health Sciences University, Brooklyn, NY 11203, USA
- Geoffrey S. Pitt
- Cardiovascular Research Institute, Weill Cornell Medicine, New York, NY 10021, USA
- Todd Evans
- Department of Surgery, Weill Cornell Medical College, New York, NY 10065, USA; Corresponding author
- Shuibing Chen
- Department of Surgery, Weill Cornell Medical College, New York, NY 10065, USA; Corresponding author
- Journal volume & issue
-
Vol. 25,
no. 4
p. 104153
Abstract
Summary: The sinoatrial node (SAN) is the primary pacemaker of the heart. The human SAN is poorly understood due to limited primary tissue access and limitations in robust in vitro derivation methods. We developed a dual SHOX2:GFP; MYH6:mCherry knockin human embryonic stem cell (hESC) reporter line, which allows the identification and purification of SAN-like cells. Using this line, we performed several rounds of chemical screens and developed an efficient strategy to generate and purify hESC-derived SAN-like cells (hESC-SAN). The derived hESC-SAN cells display molecular and electrophysiological characteristics of bona fide nodal cells, which allowed exploration of their transcriptional profile at single-cell level. In sum, our dual reporter system facilitated an effective strategy for deriving human SAN-like cells, which can potentially be used for future disease modeling and drug discovery.
