Microenvironment optimization enables kidney organoid longevity with epithelial-endothelial joint basement membrane...

Area of Science:

• Biotechnology
• Regenerative Medicine
• Developmental Biology

Background:

• Kidney organoids typically degrade in long-term cultures.
• Existing kidney organoids lack essential joint basement membranes found in renal tissue.

Purpose of the Study:

• To overcome limitations in kidney organoid culture by optimizing the microenvironment.
• To enhance organoid longevity, yield, and vascularization for improved renal tissue engineering.

Main Methods:

• Supplementing standard media with tubular-enhancing factors (TEFs) and vascular-enhancing factors (VEFs).
• Utilizing replating techniques to increase endothelial cell yield and invasiveness.
• Employing transcriptomic and imaging analyses to assess organoid structure and function over time.

Main Results:

• TEFs significantly improved organoid yield and longevity in static cultures.
• VEFs and replating enhanced endothelial cell proliferation and invasiveness.
• Organoids maintained nephron structures for six months, showing increased metabolism, signaling, differentiation, and aging pathways.
• TEFs and VEFs induced self-assembly of joint basement membranes in suspension cultures.

Conclusions:

• Microenvironment optimization enables longitudinal stabilization and advanced vascularization of kidney organoids.
• This approach provides a valuable resource for long-term kidney organoid studies and tissue engineering applications.