Department of Surgery, Hanyang University College of Medicine, Seoul, 04763, Republic of Korea; HY Indang Center of Regenerative Medicine and Stem Cell Research, Hanyang University, Seoul, 04763, Republic of Korea.
Digital Manufacturing Process Group, Korea Institute of Industrial Technology, 113-58 Seohaean-ro, Siheungsi, Gyeonggi-do, 15014, Republic of Korea.
Biomaterials. 2021 Jul;274:120899. doi: 10.1016/j.biomaterials.2021.120899. Epub 2021 May 18.
Recently, use of cell sheets with bio-applicable fabrication materials for transplantation has been an attractive approach for the treatment of patients with liver failure. However, renewable and scalable cell sources for engineered tissue patches remain limited. We previously reported a new type of proliferating bipotent human chemically derived hepatic progenitor cells (hCdHs) developed by small molecule-mediated reprogramming. Here, we developed a patient-specific hepatic cell sheet constructed from liver biopsy-derived hCdHs on a multiscale fibrous scaffold by combining electrospinning and three-dimensional printing. Analysis of biomaterial composition revealed that the high-density electrospun sheet was superior in increasing the functional properties of hCdHs. Furthermore, the hepatic patch assembled by multilayer stacking with alternate cell sheets of hCdHs and human umbilical vein endothelial cells (HUVECs) recapitulated a liver tissue-like structure, with histological and morphological shape and size similar to those of primary human hepatocytes, and exhibited a significant increase in hepatic functions such as albumin secretion and activity of cytochrome P450 during in vitro hepatic differentiation compared with that in hCdH cells cultured in a two-dimensional monolayer. Interestingly, in the hepatic patch, the induction of functional hepatocytes was associated with both the electrospun fibrous-facilitated oncostatin M signaling and selective activation of AKT signaling by HUVECs. Notably, upon transplantation into a mouse model of therapeutic liver repopulation, the hepatic patch effectively repopulated the damaged parenchyma and induced the restoration of liver function with healthy morphology in the lobe and an improved survival rate (>70%) in mice. Overall, these results suggested that liver biopsy-derived hCdHs can be an efficient alternative source for developing hepatic cell sheets and patches with potential clinical applications in tissue engineering to advance liver regeneration.
最近，使用具有生物适用性制造材料的细胞片进行移植已成为治疗肝功能衰竭患者的一种有吸引力的方法。然而，用于工程组织贴片的可再生和可扩展的细胞源仍然有限。我们之前报道了一种通过小分子介导的重编程开发的新型增殖性双潜能人化学衍生肝祖细胞（hCdHs）。在这里，我们开发了一种由肝活检衍生的 hCdHs 在多尺度纤维支架上构建的患者特异性肝细胞膜片，该支架结合了静电纺丝和三维打印。对生物材料组成的分析表明，高密度静电纺丝片在提高 hCdHs 的功能特性方面具有优势。此外，通过多层堆叠，将 hCdHs 和人脐静脉内皮细胞（HUVEC）的细胞片交替组装的肝贴片再现了类似于肝组织的结构，其组织学和形态形状和大小与人原代肝细胞相似，并且在体外肝分化过程中与在二维单层中培养的 hCdH 细胞相比，白蛋白分泌和细胞色素 P450 活性等肝功能显著增加。有趣的是，在肝贴片中，功能性肝细胞的诱导与静电纺丝纤维促进的 Oncostatin M 信号和 HUVEC 选择性激活 AKT 信号有关。值得注意的是，在将其移植到治疗性肝再灌注的小鼠模型中后，肝贴片有效地再灌注受损实质，并诱导肝脏功能恢复，肝叶形态健康，小鼠存活率提高（>70%）。总体而言，这些结果表明，肝活检衍生的 hCdHs 可以作为开发具有潜在临床应用的肝细胞膜片和贴片的有效替代来源，以推进组织工程中的肝再生。
