Institute of Reproductive and Developmental Biology, Faculty of Medicine, Imperial College London, Hammersmith Hospital Campus, Du Cane Road, London, W12 0NN, UK.
Stem Cell Res Ther. 2023 Nov 1;14(1):314. doi: 10.1186/s13287-023-03542-x.
Human pluripotent stem cell (hPSC)-derived hepatocyte-like cells (HLCs) can provide a valuable in vitro model for disease modelling and drug development. However, generating HLCs with characteristics comparable to hepatocytes in vivo is challenging. Extracellular matrix (ECM) plays an important role in supporting liver development and hepatocyte functions, but their impact on hepatocyte differentiation and maturation during hPSC differentiation remains unclear. Here, we investigate the effects of two ECM components-Matrigel and type I collagen on hepatic differentiation of human embryonic stem cells (hESCs).
hESC-derived HLCs were generated through multistage differentiation in two-dimensional (2D) and three-dimensional (3D) cultures, incorporating either type I collagen or Matrigel during hepatic specification and maturation. The resulting HLCs was characterized for their gene expression and functionality using various molecular and cellular techniques.
Our results showed that HLCs cultured with collagen exhibited a significant increase in albumin and alpha-1 anti-trypsin expression with reduced AFP compared to HLCs cultured with Matrigel. They also secreted more urea than Matrigel cultures. However, these HLCs exhibited lower CYP3A4 activity and glycogen storage than those cultured with Matrigel. These functional differences in HLCs between collagen and Matrigel cultures closely resembled the hepatocytes of periportal and pericentral zones, respectively.
Our study demonstrates that Matrigel and collagen have differential effects on the differentiation and functionality of HLCs, which resemble, to an extent, hepatic zonation in the liver lobules. Our finding has an important impact on the generation of hPSC-HLCs for biomedical and medical applications.
人类多能干细胞（hPSC）衍生的肝细胞样细胞（HLC）可为疾病建模和药物开发提供有价值的体外模型。然而，生成具有与体内肝细胞相当特征的 HLC 具有挑战性。细胞外基质（ECM）在支持肝脏发育和肝细胞功能方面起着重要作用，但它们对 hPSC 分化过程中肝细胞分化和成熟的影响尚不清楚。在这里，我们研究了两种 ECM 成分-Matrigel 和 I 型胶原对人胚胎干细胞（hESC）肝分化的影响。
通过二维（2D）和三维（3D）培养中的多阶段分化生成 hESC 衍生的 HLC，在肝特化和成熟过程中分别使用 I 型胶原或 Matrigel。使用各种分子和细胞技术对产生的 HLC 进行基因表达和功能特性的表征。
我们的结果表明，与在 Matrigel 中培养的 HLC 相比，在胶原中培养的 HLC 表现出白蛋白和α-1 抗胰蛋白酶表达的显著增加，而 AFP 减少。它们还比 Matrigel 培养物分泌更多的尿素。然而，这些 HLC 的 CYP3A4 活性和糖原储存低于在 Matrigel 中培养的 HLC。胶原和 Matrigel 培养物中 HLC 的这些功能差异与肝小叶中分别具有门脉周围和中央区特征的肝细胞非常相似。
我们的研究表明，Matrigel 和胶原对 HLC 的分化和功能具有不同的影响，在一定程度上类似于肝小叶中的肝区带化。我们的发现对用于生物医学和医学应用的 hPSC-HLC 的生成具有重要影响。
