Current methods and challenges in the comprehensive characterization of human pluripotent stem cells

Area of Science:

• Stem Cell Biology
• Regenerative Medicine
• Drug Discovery

Background:

• Pluripotent stem cells (PSCs) are vital for research, drug discovery, and regenerative medicine.
• Generating PSCs is becoming easier, but variations in origin, derivation, and culture affect cell characteristics.
• Consistent characterization is essential to confirm pluripotency, quality, identity, and safety of PSC lines.

Purpose of the Study:

• To provide a comprehensive review of current practices in PSC characterization.
• To explore challenges in selecting markers and developing scalable characterization methods.
• To discuss emerging trends, including alternative and in silico assays for pluripotency determination.

Main Methods:

• Review of current standard assays for assessing pluripotency, quality, identity, and safety of PSCs.
• Analysis of marker selection and the development of scalable methods for characterization.
• Exploration of novel approaches, including in silico assays, as alternatives or supplements to traditional methods.

Main Results:

• Current characterization relies on marker expression in undifferentiated and differentiated states for functional pluripotency.
• Challenges exist in marker selection, assay scalability, and standardization across different PSC lines.
• Emerging trends show promise for alternative assays, particularly in silico methods, to enhance PSC characterization.

Conclusions:

• Standardized and scalable characterization assays are critical for reliable use of PSCs in research and clinical applications.
• Addressing challenges in marker selection and assay development is key to advancing the field.
• Innovative approaches, such as in silico assays, offer potential for more efficient and comprehensive PSC characterization.