Reprogramming Urine-Derived Cells using Commercially Available Self-Replicative RNA and a Single Electroporation.

Area of Science:

• Stem Cell Biology
• Regenerative Medicine
• Cellular Reprogramming

Background:

• Human-induced pluripotent stem cells (hiPSCs) hold great promise for regenerative medicine and disease modeling.
• Deriving hiPSCs typically requires invasive tissue sampling or specialized cell sources.
• Urine-derived cells (UDCs) offer a non-invasive and readily accessible source for cell reprogramming.

Purpose of the Study:

• To establish and validate an efficient protocol for generating hiPSCs from adult UDCs.
• To utilize a commercially available self-replicative RNA (ReproRNA-OKSGM) for reprogramming.
• To provide guidance on pluripotency characterization of the generated hiPSC lines.

Main Methods:

• Reprogramming of UDCs using ReproRNA-OKSGM via electroporation.
• Quantification of transfection efficiency using flow cytometry for OCT3/4 expression (≥0.1%).
• Isolation and expansion of hiPSC colonies, followed by pluripotency marker analysis and differentiation assays.

Main Results:

• Successful generation of hiPSC colonies from UDCs within 3 weeks post-transfection.
• Demonstrated expression of multiple pluripotency markers in UDC-derived hiPSCs.
• UDC-derived hiPSCs exhibited successful differentiation into all three germ layers and possessed normal karyotypes.

Conclusions:

• The ReproRNA-OKSGM method provides an efficient, accessible, and straightforward approach for generating hiPSCs from UDCs.
• This protocol facilitates the derivation of patient-specific hiPSCs from a non-invasive source.
• The generated hiPSCs are functionally pluripotent and suitable for further research and therapeutic applications.