UTF1 Expression is Important for the Generation and Maintenance of Human iPSCs

Area of Science:

• Stem cell biology
• Epigenetics
• Gene regulation

Background:

• Undifferentiated embryonic cell transcription factor 1 (UTF1) is vital for embryonic development and pluripotency maintenance.
• UTF1 is highly expressed in pluripotent stem cells, but its role in human induced pluripotent stem cells (iPSCs) remains understudied.
• Previous research in murine models highlights UTF1's importance, necessitating investigation in human iPSCs.

Purpose of the Study:

• To investigate the role of UTF1 in maintaining the pluripotency and stability of human iPSCs.
• To analyze the effects of UTF1 depletion on iPSC reprogramming efficiency, cellular morphology, and pluripotency markers.
• To explore the mechanistic impact of UTF1 downregulation on iPSC differentiation and viability.

Main Methods:

• CRISPR/Cas9 gene editing was used to create UTF1 knockout in human fibroblasts and iPSCs.
• Episomal vectors facilitated the reprogramming of UTF1 knockout fibroblasts into iPSCs.
• Western blotting, PCR, and flow cytometry assessed pluripotency markers, cellular morphology, and viability.
• An shRNA was integrated to downregulate UTF1 expression for mechanistic studies.

Main Results:

• UTF1 knockout significantly reduced reprogramming efficiency and increased spontaneous differentiation in human iPSCs.
• Gradual UTF1 depletion led to altered cellular morphology and decreased expression of OCT4 and SOX2 pluripotency markers.
• Unlike complete knockout, partial UTF1 downregulation did not induce apoptosis, indicating pluripotency loss independent of apoptotic pathways.

Conclusions:

• UTF1 is essential for preserving the pluripotency and viability of human iPSCs.
• UTF1 depletion negatively impacts fundamental stem cell properties, posing challenges for therapeutic applications.
• Further research into UTF1's mechanistic pathways controlling pluripotency and differentiation is needed to enhance iPSC stability for clinical use.