On March 8, Cell Stem Cell published online the research achievements of the Huilijian team of the Center for Excellence and Innovation in Molecular Cell Science of the Chinese Academy of Sciences, entitled "Preferred efficiency and safety of encapsulated proliferating human hematopoietic organizations in treating life failure.". This study achieved the large-scale preparation of proliferative human liver cell like organs (eLO) encapsulated in sodium alginate microcapsules, and demonstrated the therapeutic effectiveness of eLO intraperitoneal transplantation in 80% liver resection induced liver failure model and drug induced acetaminophen (APAP) induced liver failure mouse model. In postoperative liver failure, eLO improves liver failure by maintaining the homeostasis of the hepatic intestinal axis, protecting the intestinal barrier, reducing endotoxins from the intestine, and promoting liver regeneration. This study validated the genomic stability of ProliHH cells and the in vivo safety of eLO transplantation.

This study identified the cell viability and liver cell maturation function of mass-produced eLO and found that the cell viability of eLO remained above 85%, comparable to ProliHH like organs (LO) that were not encapsulated in microcapsules. eLO also exhibited similar levels of mature liver gene expression, protein secretion, and metabolic activity to LO, proving that microcapsule encapsulation did not affect the liver cell function of LO maturation.

Simulating clinical postoperative liver failure and small liver syndrome, researchers constructed an 80% liver resection induced liver failure model (ePH) and injected eLO into the abdominal cavity of mice. It was found that microencapsulation can form an immune protective barrier, allowing the transplanted LO to maintain good function within 7 days and making subsequent cell recovery possible. Research has found that liver cells with 5% liver mass have significant therapeutic effects on ePH mice. Compared with the empty microcapsule treatment group, the survival rate of mice in the eLO treatment group was improved, accompanied by a decrease in liver injury biochemical indicators such as AST and ALT, and a significant recovery in liver functions such as coagulation function and blood ammonia lowering ability, as well as IL-6 and TNF- α Wait for inflammation indicators to decrease. In the liver of mice treated with eLO, researchers observed a significant increase in liver cell proliferation starting from the second day after liver resection. On the 14th day, the liver to body weight ratio returned to a similar level as normal mice, and the transcriptome of the liver of mice treated with eLO was also close to normal liver. In addition, studies have shown that eLO also has significant therapeutic effects on APAP induced liver failure in mice.

Furthermore, researchers explored the mechanism of eLO in treating ePH. Through gene expression level, protein synthesis level, and staining analysis of eLO recovered from the abdominal cavity of ePH mice, research has shown that eLO has the ability of ammonia metabolism, urea synthesis, and glycogen storage, which can improve symptoms such as high blood ammonia and low blood sugar in ePH mice. In order to investigate the promoting effect of eLO on liver regeneration, researchers found that compared with the empty microcapsule treatment group, the intestinal morphology and permeability of the eLO treatment group mice were closer to normal mice, and the serum endotoxin levels from the intestine were significantly reduced. Subsequently, the inflammatory cytokine levels in the eLO treatment group mice decreased, reducing liver inflammation and promoting liver regeneration. Due to the significant decrease in total bile acid levels after eLO treatment compared to the empty microcapsule control group mice, the accumulated bile acid can disrupt the intestinal barrier and increase intestinal permeability. Researchers fed ePH mice with significantly increased bile acid levels after liver resection to increase intestinal permeability. They found that even after eLO treatment, the survival rate of mice was significantly reduced, and the level of inflammatory factor IL-6 was significantly upregulated. These experimental data indicate that eLO therapy not only provides liver function support, but also helps liver regeneration by protecting the intestinal barrier and reducing gut derived toxins.

This study analyzed the biological distribution, toxicity, and tumorigenicity of eLO, and the results of hematological toxicity, blood biochemical indicators, and anatomical pathology tests all confirmed the safety of eLO transplantation. In vitro, the genomic stability and tumorigenicity of ProliHH cultured in vitro were studied and identified through karyotype identification and whole genome sequencing, further validating the safety of the cell.

This study achieved the large-scale preparation of proliferative human liver cell like organs encapsulated in sodium alginate microcapsules, and demonstrated the effectiveness of eLO intraperitoneal transplantation in 80% liver resection and acetaminophen induced liver failure mouse models. Meanwhile, the study validated the genomic stability of cells and the in vivo safety of eLO transplantation in mouse models, laying the foundation for clinical research on eLO.

The research work was supported by the Chinese Academy of Sciences, the National Natural Science Foundation of China, the Ministry of Science and Technology, Shanghai Municipal Commission of Science and Technology and other projects.

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