A new, economical, and easy protocol to culture 3D mouse hepatoid and cholangoid

Despite the high efficiency of self-regeneration in liver, liver stem cell organoid culture is way behind other cell types such as intestinal organoid (Enteroid). The contributing factor is the lack of effi¬cient technique to isolate liver cells and to culture liver stem cell organoid. Here we reported a new, eco¬nomical, and simple method to generate 3D hepatocyte organoid (Hepatoid) and cholangiocyte organoid (Cholangoid). We have isolated liver cells using ice-cold 3mM EDTA PBS medium. The isolated cells were cultured on 50% Conditional Medium. After 1 or 4 days post isolation and culture, Cholangoid and Hepatoid were readily to be detected under light microscope and they continued to grow well after several passages. This method could be used to isolate and culture human liver organoids. This feasible protocol will greatly facilitate more research on liver function and diseases using liver cell organoids.

Keywords: Organoid; Stem cell; 3D culture; Liver