Tissue Eng Regen Med.  2024 Jul;21(5):749-759. 10.1007/s13770-024-00632-6.

Exploring the Cocktail Factor Approach to Generate Salivary Gland Progenitors through Co-Culture Techniques

• Zhang Y
• Yan S
• Mei Z
• Zhang H
• Ding C
• Zhang S
• Wei S

• KMID: 2557869
• DOI: http://doi.org/10.1007/s13770-024-00632-6

Abstract

BACKGROUND
The derivation of salivary gland (SG) progenitors from pluripotent stem cells (PSCs) presents significant potential for developmental biology and regenerative medicine. However, the existing protocols for inducing SG include limited factors, making it challenging to mimic the in vivo microenvironment of embryonic SGs.
METHODS
We reported a cocktail factor approach to promote the differentiation of mouse embryonic stem cell (mESC)-derived oral epithelium (OE) into SG progenitors through a three-dimensional co-culture method. Upon confirming that the embryonic SG can promote the differentiation of mESC-derived OE, we performed RNA sequence analysis to identify factors involved in the differentiation of SG progenitors.
RESULTS
Our findings highlight several efficient pathways related to SG development, with frequent appearances of four factors: IFN-c, TGF-b2, EGF, and IGF-1. The combined treatment using these cocktail factors increased the expression of key SG progenitor markers, including Sox9, Sox10, Krt5, and Krt14. However, absence of any one of these cocktail factors did not facilitate differentiation. Notably, aggregates treated with the cocktail factor formed SG epitheliallike structures and pre-bud-like structures on the surface.
CONCLUSION
In conclusion, this study offers a novel approach to developing a differentiation protocol that closely mimics the in vivo microenvironment of embryonic SGs. This provides a foundation for generating PSC-derived organoids with near-physiological cell behaviors and structures.

Keyword

Mouse embryonic stem cells; Salivary glands; Organoids; Coculture techniques