Acute liver failure (ALF) is a complicated condition that is characterized by global hepatocyte death and often requires immediate liver transplantation. However, this therapy is limited by shortage of donor organs. Mesenchymal stem cells (MSCs) and hepatocytes are two attractive sources of cell-based therapies to treat ALF. The combined transplantation of hepatocytes and MSCs is considered to be more effective for the treatment of ALF than single-cell transplantation. We have previously demonstrated that HNF4α-overexpressing human umbilical cord MSCs (HNF4α-UMSCs) promoted the expression of hepatic-specific genes. In addition, microencapsulation allows exchange of nutrients, forming a protective barrier to the transplanted cells. Moreover, encapsulation of hepatocytes improves the viability and synthetic ability of hepatocytes and circumvents immune rejection. This study aimed to investigate the therapeutic effect of microencapsulation of hepatocytes and HNF4α-UMSCs in ALF mice. Human hepatocytes and UMSCs were obtained separately from liver and umbilical cord, followed by co-encapsulation and transplantation into mice by intraperitoneal injection. LPS/D-gal was used to induce ALF by intraperitoneal injection 24 h after transplantation. In addition, Raw 264.7 cells (a macrophage cell line) were used to elucidate the effect of HNF4α-UMSCs-hepatocyte microcapsules on polarization of macrophages. The protein chip was used to define the important paracrine factors in the conditioned mediums (CMs) of UMSCs and HNF4α-UMSCs and investigate the possible mechanism of HNF4α-UMSCs for the treatment of ALF in mice. HNF4α-UMSCs can enhance the function of primary hepatocytes in alginate–poly-L-lysine–alginate (APA) microcapsules. The co-encapsulation of both HNF4α-UMSCs and hepatocytes achieved better therapeutic effects in ALF mice by promoting M2 macrophage polarization and reducing inflammatory response mainly mediated by the paracrine factor HB-EGF secreted by HNF4α-UMSCs. The present study confirms that the co-encapsulation of HNF4α-UMSC and hepatocytes could exert therapeutic effect on ALF mainly by HB-EGF secreted by HNF4α-UMSCs and provides a novel strategy for the treatment of ALF.

中文翻译：

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HNF4α过表达的UMSCs和人类原代肝细胞的共包裹可改善小鼠急性肝衰竭

急性肝衰竭（ALF）是一种复杂的疾病，其特征是整体肝细胞死亡，通常需要立即进行肝移植。但是，这种疗法受到供体器官短缺的限制。间充质干细胞（MSCs）和肝细胞是治疗ALF的基于细胞疗法的两个有吸引力的来源。肝细胞和MSC的联合移植被认为比单细胞移植更有效地治疗ALF。以前我们已经证明过表达HNF4α的人脐带MSC（HNF4α-UMSCs）促进了肝特异性基因的表达。另外，微囊化允许营养物交换，从而形成对移植细胞的保护性屏障。此外，肝细胞的包囊改善了肝细胞的活力和合成能力，并规避了免疫排斥。这项研究旨在研究肝细胞和HNF4α-UMSCs微囊化对ALF小鼠的治疗作用。从肝和脐带中分别获得人肝细胞和UMSC，然后通过腹膜内注射共包囊和移植到小鼠中。移植后24小时腹腔注射LPS / D-gal诱导ALF。此外，使用Raw 264.7细胞（巨噬细胞系）来阐明HNF4α-UMSCs-肝细胞微胶囊对巨噬细胞极化的影响。该蛋白芯片用于定义UMSC和HNF4α-UMSC的条件培养基（CMs）中的重要旁分泌因子，并研究HNF4α-UMSCs治疗小鼠ALF的可能机制。HNF4α-UMSCs可增强藻酸盐-聚-L-赖氨酸-藻酸盐（APA）微胶囊中原代肝细胞的功能。HNF4α-UMSCs和肝细胞的共包封可通过促进M2巨噬细胞极化和减少主要由HNF4α-UMSC分泌的旁分泌因子HB-EGF介导的炎症反应，在ALF小鼠中获得更好的治疗效果。本研究证实，HNF4α-UMSC与肝细胞的共包被主要通过HNF4α-UMSCs分泌的HB-EGF对ALF发挥治疗作用，并为ALF的治疗提供了新的策略。HNF4α-UMSCs可增强藻酸盐-聚-L-赖氨酸-藻酸盐（APA）微胶囊中原代肝细胞的功能。HNF4α-UMSCs和肝细胞的共包封可通过促进M2巨噬细胞极化和减少主要由HNF4α-UMSC分泌的旁分泌因子HB-EGF介导的炎症反应，在ALF小鼠中获得更好的治疗效果。本研究证实，HNF4α-UMSC与肝细胞的共包被主要通过HNF4α-UMSCs分泌的HB-EGF对ALF发挥治疗作用，并为ALF的治疗提供了新的策略。HNF4α-UMSCs可增强藻酸盐-聚-L-赖氨酸-藻酸盐（APA）微胶囊中原代肝细胞的功能。HNF4α-UMSCs和肝细胞的共包封可通过促进M2巨噬细胞极化和减少主要由HNF4α-UMSC分泌的旁分泌因子HB-EGF介导的炎症反应，在ALF小鼠中获得更好的治疗效果。本研究证实，HNF4α-UMSC与肝细胞的共包被主要通过HNF4α-UMSCs分泌的HB-EGF对ALF发挥治疗作用，并为ALF的治疗提供了新的策略。HNF4α-UMSCs和肝细胞的共包封通过促进M2巨噬细胞极化和减少主要由HNF4α-UMSC分泌的旁分泌因子HB-EGF介导的炎症反应，在ALF小鼠中获得更好的治疗效果。本研究证实，HNF4α-UMSC与肝细胞的共包被主要通过HNF4α-UMSCs分泌的HB-EGF对ALF发挥治疗作用，并为ALF的治疗提供了新的策略。HNF4α-UMSCs和肝细胞的共包封可通过促进M2巨噬细胞极化和减少主要由HNF4α-UMSC分泌的旁分泌因子HB-EGF介导的炎症反应，在ALF小鼠中获得更好的治疗效果。本研究证实，HNF4α-UMSC与肝细胞的共包被主要通过HNF4α-UMSCs分泌的HB-EGF对ALF发挥治疗作用，并为ALF的治疗提供了新的策略。