ABSTRACT. The liver is a unique organ that is endowed with a plethora of specialized functions. Most of its functional traits are controlled by hepatocytes. Primary hepatocytes have been used widely in in vitro models to understand the biological processes occurring in the liver. There are a number of methods used to separate hepatocytes, but the cell activity and purity are much lower in this condition. On the basis of previous research, in this study, the two-step collagenase perfusion technique was used for isolating hepatocytes. The key proteins of hepatocytes, cytokeratin-18 (CK-18) and albumin (ALB), were used to identify cells, and their contents were evaluated by immunohistochemistry and Western blotting. The results showed that the isolated hepatocytes comprised more than 96% of the corresponding protein volume stability. Therefore, this method was demonstrated to be reliable for identifying hepatocytes.
ABSTRACT. Lactoferrin (Lf) is an iron-binding glycoprotein that is produced by mucosal epithelial cells in mammals. Lf has non-immune natural defense functions and biological functions in addition to and distinct from its role in regulating inflammatory responses. Lf also improved some physiological and immunological parameters. Lf is a biomarker for monitoring medical treatment in inflammatory bowel diseases. Current LF research focuses on iron absorption, antimicrobial activity, and the modulation of iron metabolism during inflammation.No systematic research about Lf expression levels in mouse mammary glands during pregnancy and lactation exists. We investigated Lf mRNA expression levels in mouse mammary glands by collecting samples on days 1, 6, 12, and 18 of pregnancy and lactation (six mice per group). The expression levels of Lf mRNA were measured by semi-quantitative reverse transcription polymerase chain reaction using GAPDH as an internal control. Lf mRNA was not expressed in mammary glands on days 1, 6, and 12 of pregnancy, but it was expressed on day 18 (IOD: integrated optical density; Lf IOD /GAPDH IOD = 0.46). Lf expression levels were higher during lactation stages than during pregnancy stages, and it stabilized at 0.71-0.73 (Lf IOD /GAPDH IOD ) from day 1 to 12 of lactation; however, the difference was not significant (P > 0.05). At day 18 of lactation, Lf expression began to decline (Lf IOD /GAPDH IOD = 0.61), but the difference was not significant (P > 0.05). Based on these results, the variation in Lf expression levels during developmental stages may be related to its regulatory role in mouse mammary gland immunity.
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