Towards uterus tissue engineering: a comparative study of sheep uterus decellularisation

Area of Science:

• Biomedical Engineering
• Regenerative Medicine
• Reproductive Biology

Background:

• Current uterus transplantation faces challenges including donor availability, complex surgery, immunosuppression, and rejection.
• Tissue engineering using decellularized uterine scaffolds offers a potential alternative to overcome these limitations.
• Previous rodent studies show promise, necessitating larger animal models for further investigation.

Purpose of the Study:

• To establish effective decellularization protocols for sheep uterus scaffolds.
• To prepare for future in vivo bioengineering experiments in sheep models.
• To evaluate the suitability of decellularized scaffolds for supporting uterine cells.

Main Methods:

• Three decellularization protocols were developed using vascular perfusion of sheep uteri with different solutions: 0.5% sodium dodecyl sulfate (SDS), 2% sodium deoxycholate (SDC), or sequential 2% SDC and 1% Triton X-100.
• Scaffolds were analyzed for extracellular matrix integrity, mechanical properties, and cytotoxicity.
• The ability of scaffolds to support fetal sheep stem cells in vitro was assessed.

Main Results:

• All three protocols successfully decellularized sheep uteri while preserving extracellular matrix integrity.
• The 2% sodium deoxycholate protocol (Protocol 2) yielded the most favorable results based on quantitative analysis.
• All decellularized scaffolds supported fetal sheep stem cell proliferation and maintained their undifferentiated phenotype for two weeks without cytotoxicity.

Conclusions:

• Effective decellularization protocols for sheep uteri were established, preserving crucial extracellular matrix components.
• Sodium deoxycholate-based protocols demonstrate significant potential for uterine tissue engineering.
• These validated protocols provide a foundation for future in vivo studies in larger animal models, advancing uterus bioengineering.