人類及豬主動脈之條件培養基應用於肝臟再生的可行性評估

Abstract

肝臟具有強大的再生能力，但受到外科手術副作用或慢性疾病等因素影響時，其再生功能可能受到抑制。為了在肝臟受損後迅速恢復人體代謝功能，促進肝臟再生具有重要臨床意義。在此過程中，微環境調控與血管新生扮演關鍵角色。主動脈作為富含血管相關細胞與胞外基質（extracellular matrix, ECM）之組織，其衍生之條件培養基（conditioned medium, CM）可能含有多種促進再生潛力的生物活性因子，具有應用於肝臟再生相關研究之潛力。本研究利用人類與豬主動脈製備CM，並以HepG2肝癌細胞作為體外模型，評估CM對細胞存活力、尿素合成與白蛋白分泌等肝功能指標之影響。實驗結果顯示，豬主動脈CM在低血清環境下可有效增進細胞活性與尿素生成，而白蛋白分泌則無顯著影響。人類主動脈CM則在低濃度時可支持細胞功能，但高濃度條件下可能產生細胞毒性。另將HepG2細胞直接接種於冷凍保存之人類與豬主動脈組織上，結果證實兩者皆能支持細胞附著與基本功能。這項研究顯示豬主動脈CM在提升細胞活性與代謝功能方面具有潛力，並且取得容易，為未來開發肝臟再生相關生醫材料提供一項具應用前景的替代選擇。

The liver possesses a remarkable regenerative capacity; however, this function may be impaired by factors such as surgical trauma or chronic liver disease. Promoting liver regeneration is therefore of great clinical importance to rapidly restore metabolic function following hepatic injury. During the regenerative process, microenvironmental modulation and angiogenesis play pivotal roles. The aorta, a tissue rich in vascular-related cells and extracellular matrix (ECM) components, may serve as a source of regenerative cues through its conditioned medium (CM), which contains various bioactive molecules with pro-regenerative potential. In this study, CM was prepared from human and porcine aortic tissues, and its effects on cell viability, urea synthesis, and albumin secretion were evaluated using the HepG2 hepatocellular carcinoma cell line as an in vitro model. The results demonstrated that porcine aorta-CM significantly enhanced cell viability and urea production under low-serum conditions, although no significant effect on albumin secretion was observed. Human aorta-CM supported cellular functions at low concentrations, while high concentrations induced cytotoxicity. Furthermore, HepG2 cells were directly seeded onto cryopreserved human and porcine aortic tissues, both of which were shown to support cell attachment and basic hepatic functions. Overall, this study indicates that porcine aorta-CM holds promise in enhancing cellular activity and metabolic function. Its ready accessibility and favorable bioactivity suggest porcine aorta-CM is a feasible alternative biomaterial for future liver regeneration strategies.