Yousaf Mehwish, Tayyeb Asima, Ali Gibran
National Centre of Excellence in Molecular Biology, University of the Punjab.
School of Biological Sciences, University of the Punjab, Lahore, Pakistan.
Stem Cells Cloning. 2017 Sep 20;10:21-28. doi: 10.2147/SCCAA.S139497. eCollection 2017.
Culturing of primary hepatocytes and stem cell-derived hepatocytes faces a major issue of dedifferentiation due to absence of cell-cell adhesion and 3D structures. One of the possible ways to eliminate the problem of dedifferentiation is mimicking the expression pattern of adhesion proteins during the normal developmental process of liver cells. The purpose of this study was to evaluate the expression pattern of some key adhesion proteins, namely, E-cadherin, N-cadherin, epithelial CAM (EpCAM), intracellular CAM (ICAM), collagen 1α1, α-actinin, β-catenin and vimentin, in the liver tissue during prenatal and postnatal stages. Furthermore, differences in their expression between prenatal, early postnatal and adult stages were highlighted. Wistar rats were used to isolate livers at prenatal Day 14 and 17 as well as on postnatal Day 1, 3, 7 and 14. The liver from adult rats was used as control. Both conventional and real-time quantitative polymerase chain reactions (PCRs) were performed. For most of the adhesion proteins such as E-cadherin, N-cadherin, EpCAM, ICAM, collagen 1α1 and α-actinin, low expression was observed around prenatal Day 14 and an increasing expression was observed in the postnatal period. Moreover, β-catenin and vimentin showed higher expression in the early prenatal period, which decreased gradually in the postnatal period, but still this low expression was considerably higher than that in the adult control rats. This basic knowledge of the regulation of expression of adhesion proteins during different developmental stages indicates their vital role in liver development. This pattern can be further studied and imitated under in vitro conditions to achieve better cell-cell interactions.
由于缺乏细胞间黏附及三维结构，原代肝细胞和干细胞来源的肝细胞培养面临着去分化这一主要问题。消除去分化问题的一种可能方法是模拟肝细胞正常发育过程中黏附蛋白的表达模式。本研究的目的是评估一些关键黏附蛋白，即E-钙黏蛋白、N-钙黏蛋白、上皮细胞黏附分子（EpCAM）、细胞内黏附分子（ICAM）、胶原蛋白1α1、α-辅肌动蛋白、β-连环蛋白和波形蛋白在产前和产后阶段肝脏组织中的表达模式。此外，还突出了它们在产前、产后早期和成年阶段表达的差异。使用Wistar大鼠在产前第14天和17天以及产后第1天、3天、7天和14天分离肝脏。成年大鼠的肝脏用作对照。进行了传统和实时定量聚合酶链反应（PCR）。对于大多数黏附蛋白，如E-钙黏蛋白、N-钙黏蛋白、EpCAM、ICAM、胶原蛋白1α1和α-辅肌动蛋白，在产前第14天左右观察到低表达，在产后阶段观察到表达增加。此外，β-连环蛋白和波形蛋白在产前早期表达较高，在产后阶段逐渐降低，但这种低表达仍明显高于成年对照大鼠。不同发育阶段黏附蛋白表达调控的这一基础知识表明它们在肝脏发育中起着至关重要的作用。这种模式可以在体外条件下进一步研究和模仿，以实现更好的细胞间相互作用。
