Effects of Co-Culture Media on Hepatic Differentiation of hiPSC with or without HUVEC Co-Culture

International Journal of Molecular Sciences

Leila Amini¹  Selina Greuel¹  Katrin Zeilinger¹  Fanny Knöspel¹  Nadja Strahl¹  Nora Freyer¹  Mario Monshouwer²  Frank Jacobs² 

[1] Berlin-Brandenburg Center for Regenerative Therapies (BCRT), Charité—Universitätsmedizin Berlin, Campus Virchow-Klinikum, 13353 Berlin, Germany;Janssen Research and Development, 2340 Beerse, Belgium;

关键词: human induced pluripotent stem cells (hiPSC);    hepatic differentiation;    human umbilical vein endothelial cells (HUVEC);    co-culture;   

DOI  :  10.3390/ijms18081724

来源: DOAJ

【 摘 要 】

The derivation of hepatocytes from human induced pluripotent stem cells (hiPSC) is of great interest for applications in pharmacological research. However, full maturation of hiPSC-derived hepatocytes has not yet been achieved in vitro. To improve hepatic differentiation, co-cultivation of hiPSC with human umbilical vein endothelial cells (HUVEC) during hepatic differentiation was investigated in this study. In the first step, different culture media variations based on hepatocyte culture medium (HCM) were tested in HUVEC mono-cultures to establish a suitable culture medium for co-culture experiments. Based on the results, two media variants were selected to differentiate hiPSC-derived definitive endodermal (DE) cells into mature hepatocytes with or without HUVEC addition. DE cells differentiated in mono-cultures in the presence of those media variants showed a significant increase (p < 0.05) in secretion of α-fetoprotein and in activities of cytochrome P450 (CYP) isoenzymes CYP2B6 and CYP3A4 as compared with cells differentiated in unmodified HCM used as control. Co-cultivation with HUVEC did not further improve the differentiation outcome. Thus, it can be concluded that the effect of the used medium outweighed the effect of HUVEC co-culture, emphasizing the importance of the culture medium composition for hiPSC differentiation.

【 授权许可】

Unknown