Urine-derived stem cells serve as a robust platform for generating native or engineered extracellular vesicles.

Area of Science:

• Cell Biology
• Regenerative Medicine
• Biotechnology

Background:

• Mesenchymal stromal cell (MSC) therapy shows promise but faces challenges in efficacy and safety.
• Extracellular vesicles (EVs) derived from MSCs are key mediators of therapeutic effects, but large-scale production is hindered by current methods.
• Existing MSC sources like adipose tissue and bone marrow require invasive procedures, and efficient genetic engineering of MSCs is difficult.

Purpose of the Study:

• To investigate urine-derived stem cells (USCs) as a non-invasive and abundant source for MSCs and subsequent EV production.
• To assess the feasibility of using USCs for generating both native and genetically engineered EVs.
• To evaluate the impact of donor sex and sampling time on USC characteristics and EV production potential.

Main Methods:

• USCs were isolated, expanded, and characterized from healthy donors, assessing proliferation, viability, and surface marker expression (CD73, CD90, CD105) via flow cytometry.
• EVs were produced using USCs cultured in chemically defined media under serum-free conditions.
• Genetic engineering of USCs was performed using PEI/DNA transfection with transposon technology.

Main Results:

• USCs exhibited MSC immunophenotypic characteristics, with no significant differences in clone number, doubling time, or viability across different sampling times or between sexes (though males had reduced CD73+ cells).
• USCs maintained viability and produced EVs when cultured in serum-free conditions for 72 hours.
• High-efficiency transfection of USCs was achieved, enabling the establishment of pure engineered cell cultures after selection.

Conclusions:

• USCs represent a non-invasive, robust, and readily available source for MSCs, suitable for large-scale EV production.
• The isolation and expansion of USCs are not significantly affected by donor sex or sampling time.
• USCs serve as a versatile platform for producing both native and engineered extracellular vesicles for therapeutic applications.