Long-term in vivo chimeric cells tracking in non-human primate

Area of Science:

• Stem Cell Biology
• Primate Models
• Biotechnology

Background:

• Non-human primates (NHPs) are crucial for preclinical trials but pose ethical and cost challenges.
• Developing and maintaining primate pluripotent stem cells (PSCs) in a naïve state and detecting them in vivo are significant hurdles.
• Limitations in current NHP stem cell technology restrict broader biotechnology applications.

Purpose of the Study:

• To establish a chemically defined, xeno-free culture system for primate PSCs.
• To enable in vivo tracking of engineered primate PSCs for biomedical applications.
• To explore the utility of chimeric monkey models derived from engineered PSCs.

Main Methods:

• Developed a chemically defined, xeno-free culture system for monkey PSCs.
• Characterized PSCs using gene expression and genome-wide hypomethylation analysis.
• Integrated bioluminescent reporter genes into monkey PSCs for in vivo and in vitro tracking.
• Generated chimeric monkeys with bioluminescent PSCs to track cell integration and development.

Main Results:

• Established a robust in vitro culture system for monkey PSCs, maintaining their naïve state.
• Demonstrated distinct global gene expression and hypomethylation patterns in naïve PSCs compared to primed cells.
• Successfully tracked bioluminescent PSCs in chimeric embryos and in a living chimeric monkey for over two years.
• Confirmed that engineered PSCs retained embryonic and extra-embryonic developmental potential.

Conclusions:

• The developed culture system and tracking method overcome key limitations in primate stem cell research.
• Engineered primate PSCs can be effectively tracked in vivo, facilitating the study of cell integration and development.
• This technology holds significant promise for advancing primate stem cell engineering and the use of chimeric monkey models in clinical studies.