A PLURIPOTENT STEM CELL PLATFORM FOR IN VITRO SYSTEMS GENETICS STUDIES OF MOUSE DEVELOPMENT

Area of Science:

• Stem cell biology
• Genetics
• Developmental biology

Background:

• Pluripotent stem cells (PSCs) from diverse individuals offer insights into genetic variation's impact on development.
• Understanding genetic contributions to cell and tissue development is crucial.

Purpose of the Study:

• To establish PSC lines and methods for modeling embryonic development in genetically diverse mice.
• To characterize the role of genetic variation in gene regulation and differentiation.
• To develop scalable approaches for genetic screens.

Main Methods:

• Generation and maintenance of pluripotent stem cell (PSC) lines, including epiblast stem cells (EpiSCs).
• Creation of a genetic reference panel of Diversity Outbred PSCs (n=230) using pooled in vitro fertilization.
• Development of pooled culture techniques for EpiSCs ('cell villages').

Main Results:

• Demonstrated stable maintenance of primed pluripotent stem cells (epiblast stem cells - EpiSCs) across diverse genetic backgrounds.
• Quantified the impact of genetic variation on gene regulation and directed differentiation.
• Established pooled EpiSC culture as a feasible method for large-scale genetic screens.

Conclusions:

• New PSC lines and experimental approaches enable the study of natural genetic variation in embryonic development.
• Pooled culture of Diversity Outbred EpiSCs facilitates efficient forward genetic screens.
• This work complements human genetics and stem cell biology efforts to model genetic variation and disease risk.