Hepatic differentiation of mouse bone marrow-derived mesenchymal stem cells using a novel 3D culture system

The development of novel culture systems that mimic the in vivo microenvironment may be beneficial for inducing the differentiation of stem cells and promoting liver function. In the present study, spheroid cultures and decellularized liver scaffolds (DLSs) were utilized to obtain differentiated hepatocyte‑like cells. Mouse bone marrow (BM)‑derived mesenchymal stem cells (MSCs) self‑aggregated into spheroids under low‑attachment conditions and implanted into the DLSs via a negative pressure suction device. The Albp‑ZsGreen adenoviral vector was utilized for real‑time monitoring of hepatocyte‑like cell differentiation. To detect the differentiation stages of the MSCs, immunostaining of hepatocyte markers and functional analysis was performed. Compared with traditional 2D monolayer induction, mouse BM‑MSCs spheroids and DLSs in 3D culture generated greater yields of mature, differentiated hepatocytes. In conclusion, this 3D culture system may provide a strategy for generating hepatocyte‑like cells for portable liver micro‑organs, and aid clinical hepatocyte transplantation and liver tissue engineering.