Effects of Co-Culture Media on Hepatic Differentiation of hiPSC with or without HUVEC Co-Culture
Effects of Co-Culture Media on Hepatic Differentiation of hiPSC with or without HUVEC Co-Culture
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DOI：
10.3390/ijms18081724
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发表时间：
2017-08-01
影响因子：
5.6
通讯作者：
Zeilinger, Katrin
中科院分区：
文献类型：
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作者：
Freyer, Nora;Greuel, Selina;Zeilinger, Katrin
从人诱导多能干细胞（hiPSC）中提取肝细胞在药理学研究中具有重要的应用价值。然而，hipsc源性肝细胞的完全成熟尚未在体外实现。为了促进肝分化，本研究研究了hiPSC与人脐静脉内皮细胞（HUVEC）在肝分化过程中的共培养。第一步，以肝细胞培养基（HCM）为基础，在HUVEC单培养中测试不同培养基的变化，以确定适合共培养实验的培养基。根据结果，选择两种培养基变体将hipsc衍生的终末内胚层（DE）细胞分化为添加或不添加HUVEC的成熟肝细胞。与未加修饰的培养基相比，在培养基中分化的DE细胞-胎蛋白分泌和细胞色素P450 （CYP）同工酶CYP2B6和CYP3A4活性显著增加（p < 0.05）。与HUVEC共培养不能进一步改善分化效果。由此可见，使用培养基的效果大于HUVEC共培养的效果，强调了培养基组成对hiPSC分化的重要性。
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 -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.