A stable chimeric fibroblast growth factor (FGF) can successfully replace basic FGF in human pluripotent stem cell...

Area of Science:

• Stem cell biology
• Protein engineering
• Biochemistry

Background:

• Fibroblast growth factors (FGFs) are crucial for self-renewal in human pluripotent stem cells.
• Conventional culture uses basic FGF (FGF2), which is unstable and requires frequent replenishment.
• This instability poses challenges for consistent and efficient pluripotent stem cell culture.

Purpose of the Study:

• To evaluate a heat-stable chimeric FGF variant, FGFC, as an alternative to FGF2 for maintaining human pluripotent stem cells.
• To assess the stability and efficacy of FGFC in supporting stem cell self-renewal and pluripotency.

Main Methods:

• Development of FGFC, a chimeric protein combining human FGF1 and FGF2 domains for enhanced stability.
• Culture of human embryonic stem cells and induced pluripotent stem cells using FGFC in standard medium.
• Comparative analysis of FGFC-cultured cells versus FGF2-cultured cells.

Main Results:

• FGFC demonstrated superior thermal stability and protease resistance compared to FGF1 and FGF2.
• Human pluripotent stem cells maintained with FGFC showed no significant differences in pluripotency markers, gene expression, karyotype, or differentiation potential compared to FGF2-cultured cells.
• FGFC successfully supported the self-renewal of both human embryonic stem cells and induced pluripotent stem cells.

Conclusions:

• FGFC is a stable and effective alternative to FGF2 for the maintenance of human pluripotent stem cells.
• The use of FGFC can simplify stem cell culture protocols by reducing the need for frequent growth factor addition.
• This finding has implications for advancing stem cell research and therapeutic applications.