Expanded potential stem cell media as a tool to study human developmental hematopoiesis in vitro

Area of Science:

• Stem cell biology
• Developmental biology
• Hematology

Background:

• Pluripotent stem cells (PSCs) are crucial for studying mammalian development and regenerative medicine.
• In vitro hematopoiesis from PSCs offers insights into blood formation and potential cell therapies.
• Existing culture protocols for mouse and human PSCs differ, hindering research translation.

Purpose of the Study:

• To introduce expanded potential stem cell medium (EPSCM) for maintaining human PSCs.
• To demonstrate EPSCM's utility in facilitating in vitro hematopoiesis from human PSCs.
• To showcase EPSCM's support for gene targeting in PSCs.

Main Methods:

• Human PSCs were maintained using EPSCM.
• EPSCM-maintained human PSCs underwent spontaneous embryoid body formation and in vitro differentiation.
• RNA-sequencing (RNA-seq) was used to analyze hematopoietic cell populations.
• Gene targeting via homologous recombination was performed in EPSCM.

Main Results:

• EPSCM effectively maintains human PSCs in a stable state.
• Human PSCs in EPSCM spontaneously differentiate into embryoid bodies and undergo hematopoiesis.
• At least two distinct hematopoietic cell populations were generated, confirmed by RNA-seq.
• EPSCM facilitated the generation of an SPI1 (PU.1) reporter PSC line.

Conclusions:

• EPSCM is a versatile medium for maintaining human PSCs and supporting in vitro hematopoiesis.
• This method simplifies the study of blood cell development and lineage commitment.
• EPSCM offers a valuable tool for regenerative medicine applications and fundamental research.