Establishing extended pluripotent stem cells from human urine cells

Area of Science:

• Stem Cell Biology
• Developmental Biology
• Regenerative Medicine

Background:

• Extended pluripotent stem cells (EPSCs) are crucial for research and industry due to their potential to form embryonic and extraembryonic tissues.
• Current methods for generating human EPSCs from somatic cells are inefficient and complex.

Purpose of the Study:

• To establish a novel, efficient, and feeder-free method for generating EPSCs from human induced pluripotent stem cells (iPSCs).
• To develop a defined and optimized culture medium for EPSC generation and maintenance.

Main Methods:

• Development of a new culture medium, OCM175, with optimized L-selenium-methylcysteine and ROCK inhibitors.
• Utilization of Matrigel or laminin combinations (511:521) to eliminate the need for feeder cells.
• Conversion of integration-free iPSCs derived from human urine-derived cells (hUC-iPSCs) into EPSCs using OCM175 medium.

Main Results:

• Successfully generated EPSCs (O-IPSCs) from hUC-iPSCs using the OCM175 medium in a feeder-free system.
• Demonstrated that O-IPSCs exhibit robust intra- and extra-embryonic chimerism.
• Confirmed the potential of O-IPSCs to differentiate into trophoblast ectoderm and the three primary germ layers.

Conclusions:

• The novel OCM175 medium provides a defined and optimized formulation for efficient, feeder-free generation of EPSCs.
• The O-IPSCs possess significant potential for contributing to both embryonic and extraembryonic lineages.
• This system offers a promising foundation for advancing the application of EPSCs in regenerative medicine.