Liau Ling Ling, Makpol Suzana, Azurah Abdul Ghani Nur, Chua Kien Hui
Department of Physiology, Faculty of Medicine, Universiti Kebangsaan Malaysia, Jalan Yaacob Latif, 56000, Kuala Lumpur, Malaysia.
Department of Biochemistry, Faculty of Medicine, Universiti Kebangsaan Malaysia, Jalan Yaacob Latif, 56000, Kuala Lumpur, Malaysia.
Cytotechnology. 2018 Aug;70(4):1221-1233. doi: 10.1007/s10616-018-0214-8. Epub 2018 Mar 16.
Currently, orthotopic liver transplantation is the gold standard therapy for liver failure. However, it is limited by the insufficient organ donor and risk of immune rejection. Stem cell therapy is a promising alternative treatment for liver failure. One of the most ideal sources of stem cells for regenerative medicine is adipose-derived stem cells (ADSCs). In this study, primary ADSCs seeded on cell culture insert were indirectly co-cultured with injured HepG2 to elucidate the role of ADSCs in promoting the recovery of injured HepG2 in non-contact manner. HepG2 recovery was determined by the surface area covered by cells and growth factor concentration was measured to identify the factors involved in regeneration. Besides, HepG2 were collected for q-PCR analysis of injury, hepatocyte functional and regenerative markers expression. For the ADSCs, expression of hepatogenic differentiation genes was analyzed. Results showed that non-contact co-culture with ADSCs helped the recovery of injured HepG2. ELISA quantification revealed that ADSCs secreted higher amount of HGF and VEGF to help the recovery of injured HepG2. Furthermore, HepG2 co-cultured with ADSCs expressed significantly lower injury markers as well as significantly higher regenerative and functional markers compared to the control HepG2. ADSCs co-cultured with injured HepG2 expressed significantly higher hepatic related genes compared to the control ADSCs. In conclusion, ADSCs promote recovery of injured HepG2 via secretion of HGF and VEGF. In addition, co-cultured ADSCs showed early sign of hepatogenic differentiation in response to the factors released or secreted by the injured HepG2.
目前，原位肝移植是治疗肝衰竭的金标准疗法。然而，它受到器官供体不足和免疫排斥风险的限制。干细胞疗法是一种有前景的肝衰竭替代治疗方法。脂肪来源干细胞（ADSCs）是再生医学中最理想的干细胞来源之一。在本研究中，接种在细胞培养插入物上的原代ADSCs与受损的HepG2间接共培养，以阐明ADSCs在以非接触方式促进受损HepG2恢复中的作用。通过细胞覆盖的表面积来确定HepG2的恢复情况，并测量生长因子浓度以鉴定参与再生的因子。此外，收集HepG2进行损伤、肝细胞功能和再生标志物表达的q-PCR分析。对于ADSCs，分析其肝源性分化基因的表达。结果表明，与ADSCs非接触共培养有助于受损HepG2的恢复。ELISA定量分析显示，ADSCs分泌更高量的HGF和VEGF以帮助受损HepG2的恢复。此外，与对照组HepG2相比，与ADSCs共培养的HepG2表达的损伤标志物显著更低，而再生和功能标志物显著更高。与受损HepG2共培养的ADSCs与对照ADSCs相比，表达显著更高的肝脏相关基因。总之，ADSCs通过分泌HGF和VEGF促进受损HepG2的恢复。此外，共培养的ADSCs对受损HepG2释放或分泌的因子有反应，显示出肝源性分化的早期迹象。
