Generation of Donor-Specific iPSC for Modelling Lysosomal Storage Disorders

Area of Science:

• Biotechnology
• Stem Cell Biology
• Genetics

Background:

• Induced pluripotent stem cells (iPSCs) offer powerful human cell-based models for studying genetic disorders.
• Lysosomal storage disorders (LSDs) are a group of rare inherited metabolic diseases.
• Existing models may not fully recapitulate disease pathology or be suitable for therapeutic development.

Purpose of the Study:

• To provide a detailed protocol for generating iPSCs from individuals with lysosomal storage disorders.
• To establish disease-relevant iPSC lines for drug discovery and pre-clinical testing of novel therapies.
• To demonstrate the utility of generated iPSCs for creating isogenic disease models using CRISPR/Cas technology.

Main Methods:

• Generation of iPSCs from patient-derived skin fibroblasts using episomal plasmid transfection.
• Identification and isolation of iPSC colonies via TRA-1-60 immunofluorescence.
• Quality control including pluripotency marker expression, germ layer differentiation potential, and genetic variant confirmation.

Main Results:

• Successful generation of iPSCs from a CLN2 disease donor.
• Demonstrated pluripotency and differentiation capacity of the generated iPSCs.
• Confirmation of the disease-causing genetic variant in the iPSC lines.

Conclusions:

• The protocol effectively generates patient-specific iPSCs for lysosomal storage disorders.
• These iPSCs serve as valuable tools for disease modeling and therapeutic research.
• The generated iPSCs can be utilized with gene-editing technologies to create precise isogenic models for further study.