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Updated: Dec 23, 2025
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Introducing Point Mutations into Human Pluripotent Stem Cells Using Seamless Genome Editing
Published on: May 10, 2020
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Improving single-cell cloning workflow for gene editing in human pluripotent stem cells.
Yi-Hsien Chen1, Shondra M Pruett-Miller2
1Washington University School of Medicine, Department of Genetics, St. Louis 63110, USA; Genome Engineering and iPSC Center, USA.
Stem Cell Research
|August 13, 2018
Summary
This study presents a new workflow for culturing single human pluripotent stem cells (hPSCs) after genome editing. The method ensures cell viability and genetic stability, enabling high-throughput applications in research and medicine.
Area of Science:
- Stem Cell Biology
- Genome Engineering
- Biotechnology
Background:
- Human pluripotent stem cells (hPSCs) are crucial for research and drug discovery.
- Genome editing of hPSCs faces challenges with cell viability and single-cell culture.
Purpose of the Study:
- To develop a robust workflow for single-cell cloning and expansion of genome-edited hPSCs.
- To overcome the limitations of harsh genome editing procedures on hPSC viability.
Main Methods:
- Utilized Stem-Flex and RevitaCell supplement with Geltrex or Vitronectin (VN).
- Implemented a feeder-free, defined single-cell culture system post-genome editing and sorting.
- Established a workflow supporting single-cell cloning and expansion.
Main Results:
- Demonstrated reliable single-cell growth of hPSCs in a defined, feeder-free environment.
- Confirmed retention of pluripotency and normal karyotype in genome-edited clones.
- Developed a time-efficient and simplified culture protocol.
Conclusions:
- The described workflow effectively supports single-cell culture of genome-edited hPSCs.
- This method enhances hPSC viability and maintains genomic integrity.
- The protocol is valuable for high-throughput hPSC culture in basic research and clinical applications.
