Immortalization of chicken embryonic liver-derived cell line by stable expression of hMRP18S-2 for serotype 4 fowl adenovirus propagation

Feng, Lei; Chen, Li; Yun, Junwen; Bi, Zhixiang; Tang, Yinghua; Wu, Peipei; Hou, Jingbo

doi:10.1016/j.biologicals.2018.04.002

Cited by 3 publications

(2 citation statements)

References 37 publications

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“…However, the mechanism via which S18 immortalizes and transforms cells remains unexplored. Recently, other researchers have used S18-2 as a tool to immortalize chicken embryonic liver cells ( 6 ).…”

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confidence: 99%

Cell stemness is maintained upon concurrent expression of RB and the mitochondrial ribosomal protein S18-2

Mushtaq

Kovalevska

Darekar

et al. 2020

Proc. Natl. Acad. Sci. U.S.A.

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Stemness encompasses the capability of a cell for self-renewal and differentiation. The stem cell maintains a balance between proliferation, quiescence, and regeneration via interactions with the microenvironment. Previously, we showed that ectopic expression of the mitochondrial ribosomal protein S18-2 (MRPS18-2) led to immortalization of primary fibroblasts, accompanied by induction of an embryonic stem cell (ESC) phenotype. Moreover, we demonstrated interaction between S18-2 and the retinoblastoma-associated protein (RB) and hypothesized that the simultaneous expression of RB and S18-2 is essential for maintaining cell stemness. Here, we experimentally investigated the role of S18-2 in cell stemness and differentiation. Concurrent expression of RB and S18-2 resulted in immortalization ofRb1−/−primary mouse embryonic fibroblasts and in aggressive tumor growth in severe combined immunodeficiency mice. These cells, which express both RB and S18-2 at high levels, exhibited the potential to differentiate into various lineages in vitro, including osteogenic, chondrogenic, and adipogenic lineages. Mechanistically, S18-2 formed a multimeric protein complex with prohibitin and the ring finger protein 2 (RNF2). This molecular complex increased the monoubiquitination of histone H2ALys119, a characteristic trait of ESCs, by enhanced E3-ligase activity of RNF2. Furthermore, we found enrichment of KLF4 at theS18-2promoter region and that theS18-2expression is positively correlated withKLF4levels. Importantly, knockdown of S18-2 in zebrafish larvae led to embryonic lethality. Collectively, our findings suggest an important role for S18-2 in cell stemness and differentiation and potentially also in cancerogenesis.

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confidence: 99%

Cell stemness is maintained upon concurrent expression of RB and the mitochondrial ribosomal protein S18-2

Mushtaq

Kovalevska

Darekar

et al. 2020

Proc. Natl. Acad. Sci. U.S.A.

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“…The cell population reached a growth of 6x10 5 cells [19]. Feng et al highlighted the utility of chicken liver cells for cultivating four types of adenoviruses [20]. Li et al suggested the use of liver cells, particularly hepatocytes, as a suitable source with high viral receptor expression for propagating ALV-J (avian leukosis virus) [21].…”

Section: Discussionmentioning

confidence: 99%

Production of Hepatocytes Derived from Chicken Embryos Across Three Developmental Stages

farzaneh,

Azizidoost

2024

Preprint

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Background: Chicken hepatocytes provide a valuable resource for conducting cell-based assays and gaining insights into the underlying mechanisms of diseases. Objective: In this study, we examined in vitro generation of chicken hepatocytes from three developmental stages. Methods: In this in vitro study, chicken embryos aged 5, 7, and 10 days were used. The proliferation of chicken hepatocytes was assessed in DMEM/F12+10% FBS medium. After 3 days, the proliferation rate of hepatocytes, hepatocytes-specific gene expression, hepatocytes-specific enzymes (ALT and AST), albumin production and urea secretion were assessed. Findings: Morphological examination of individual hepatocytes exhibited a characteristic hexagonal structure with prominent nuclei and nucleoli, surrounded by a marginal cytoplasm. E10 embryos exhibiting a markedly higher proliferation rate in comparison to those from E5 and E7 stages. In contrast to E10, approximately 50% of hepatocytes derived from E5 and E7 embryos exhibit reduced proliferation after 3 days. In the suspension culture, hepatocytes had the ability to form spheroids or hepatospheres. The expression of hepatocyte-specific genes such as AFP, ALP, FOXA2, and CYP3A4 was markedly higher in E10-derived hepatocytes compared to E5-derived hepatocytes. Conversely, the expression of CXCR4, OCT4, NANOG, and SOX17 was significantly lower in E10-derived hepatocytes than in E5-derived hepatocytes. There was a significant increase in the expression of ALT and AST in E10-derived hepatocytes compared to E5-derived hepatocytes. Furthermore, Albumin production was significantly lower in E10-derived hepatocytes compared to E5-derived hepatocytes. While, urea secretion was higher in E10-derived hepatocytes compared to E5-derived hepatocytes Conclusion: Based on our findings, we have determined that E10 represents an optimal stage for the derivation and proliferation of chicken hepatocytes in vitro.

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Enhanced productivity and stability of PRV in recombinant ST-Tret1 cells

Bao

Chen

et al. 2023

Biologicals

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Immortalization of chicken embryonic liver-derived cell line by stable expression of hMRP18S-2 for serotype 4 fowl adenovirus propagation

Cited by 3 publications

References 37 publications

Cell stemness is maintained upon concurrent expression of RB and the mitochondrial ribosomal protein S18-2

Cell stemness is maintained upon concurrent expression of RB and the mitochondrial ribosomal protein S18-2

Production of Hepatocytes Derived from Chicken Embryos Across Three Developmental Stages

Enhanced productivity and stability of PRV in recombinant ST-Tret1 cells

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