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Merck

Protocol for In Vitro Establishment and Long-Term Culture of Mouse Vaginal Organoids.

STAR protocols (2020-10-29)
Ayesha Ali, Shafiq M Syed, Pradeep S Tanwar
ABSTRACT

Healthy vaginal epithelium is essential for normal reproductive functions and protects against infectious diseases. Here, we present a protocol for developing mouse vaginal organoids from single epithelial cells. These organoids recapitulate both functional and structural characteristics of vagina in situ. This model is a powerful tool for investigating how vaginal microbiome or chemicals in contraceptives and personal hygiene products interact with stem cells and alter the epithelial dynamics, which will lead to new insights into the pathogenesis of vaginal diseases. For complete details on the use and execution of this protocol, please refer to Ali et al. (2020).

MATERIALI
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Roche
DNasi I, grade II, from bovine pancreas
Sigma-Aldrich
Trypan Blue, 0.4%, liquid, sterile-filtered, suitable for cell culture
Sigma-Aldrich
Terreno Eagle modificato di Dulbecco/miscela nutritiva Ham F-12, With L-glutamine, 15 mM HEPES, and sodium bicarbonate, liquid, sterile-filtered, suitable for cell culture
Sigma-Aldrich
EGF from mouse, recombinant, expressed in E. coli, ≥98% (SDS-PAGE), ≥98% (HPLC), suitable for cell culture