A simple method for isolation and culture of primary hepatocytes from Salvelinus leucomaenis (White-spotted Charr)

White-spotted charr (Salvelinus leucomaenis, S. I.) is adjusted anadromous fish cold water, distributed in the Far East. We have previously reported the complete mitochondrial DNA sequence of white-spotted characters (l. S. imbrius and S. l. Pluvius) in Japan.

In general, fish hepatocytes useful for the study of cellular and biochemical fish. In this study, we isolated hepatocytes from the liver and white speckled charr and use basic methods, such as digestive enzymes and low centrifugation, to analyze the molecular mechanisms involved in specific cellular responses. isolated hepatocytes can be cultured at 5-20 ° C but not 37 ° C. The morphology of hepatocytes has been altered in a manner dependent on the temperature.

The properties of hepatocytes similar to the fish live. In addition, the rate of proliferation and isolated hepatocytes damage depends on the concentration of fetal bovine serum in the culture medium. Taken together, these studies show that this simple method for the isolation and culture of hepatocytes from white speckled charr may be useful for the study of cellular biochemistry and others.

A simple method for isolation and culture of primary hepatocytes from Salvelinus leucomaenis (White-spotted Charr)
A simple method for isolation and culture of primary hepatocytes from Salvelinus leucomaenis (White-spotted Charr)

A 3D Cell Culture Model Identifies the Wnt / β-Catenin Mediated Inhibition of p53 as Critical Steps for Human Hepatocyte Regeneration

The liver is the organ that is regenerative. While the mature hepatocytes in a state of homeostasis largely silent, after injury, they quickly enter the cell cycle to restore damaged tissue.

In rodents, various models of injury has provided important insights into the fundamentals of molecular regulate hepatocyte proliferation activation silent. However, little is known about the molecular mechanisms of regeneration of human hepatocytes and experimental methods to expand primary human hepatocytes (PHH). Here, the 3D model spheroid of PHH established for the regeneration of hepatocytes and integrative studies of time-lapse multi-omics analysis showed that after the isolation of indigenous people acquire PHH liver regenerative phenotype, as seen in vivo in partial hepatectomy.

However, the proliferation is limited. By analyzing global patterns promoter activity, it is estimated that the activation of Wnt / β-catenin signaling and inhibition of p53 is an important factor that is required for the proliferation of human hepatocytes. functional validation revealed that the activation of Wnt signaling through external cues alone is sufficient to inhibit p53 and proliferation-inducing aging its target PAI1 (SERPINE1) and drive the proliferation of> 50% of all PHH.

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A scalable 3D culture model was established to study the molecular and cellular biology of human hepatocyte regeneration. Using this model, the important role of Wnt / β-catenin signaling during regeneration of human hepatocytes identified.