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Culture and Application of Human Umbilical Vein Endothelial Cells (HUVECs)

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Endothelial cells (ECs) cover the lumen of blood vessels. Human umbilical vein endothelial cells (HUVECs) are a relatively simple and available human EC type, suitable for large scale preparation [1]. Thus, HUVECs are widely used  in vitro  model for ECs to study vascular endothelium properties and the main biological pathways involved in endothelium function. Isolation and culture of HUVECs    The primary isolated  Human Umbilical Vein Endothelial Cells  are the most popular ECs used in a number of researches probably because human umbilical veins are relatively more available than other types of blood vessels. Generally, primary HUVECs need to be isolated from human umbilical vein vascular wall by a collagenase treatment, then inoculated on coated culture flask and incubated in a 37 °C-5% CO2 incubator by standard protocol with a relatively minimal equipment. Cell confluency can be monitored by phase-contrast microscopy [1]. Of course, the primary Human Umbilical Vein Endothelial Cel

A Brief Overview of Human Primary Cells

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Human Primary Cells Human primary cells  are those that have been obtained directly from the healthy or pathological  in vivo  tissue of a human subject through a mechanical or enzymatic process. They are distinct from  animal primary cells , which are commonly taken from rats, mice, dogs, horses, or pigs; and  immortalized cell lines , which, through spontaneous mutation or deliberate modification, can be passaged  in vitro  indefinitely. These primary cells are widely used in biochemistry research to study metabolic processes, cell signaling, and related physiological phenomena. They can be isolated from many kinds of tissues, including those in the skin, blood, kidneys, bone marrow, musculoskeletal system, and cardiovascular system.   In their utility for various research applications, human primary cells share much in common with immortalized cell lines. Unlike cell lines, however, primary cells have a limited lifespan. Yet human primary cells also confer several research advantage

Luciferase reporter genes: How to use them in molecular biology?

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  Bioluminescence  is a powerful imaging modality for monitoring molecular and cellular features in real time [1].   Luciferases , known as bioluminescent enzymes, can catalyze the oxidation of small molecule substrates, such as luciferins, to release visible light. Currently, the luciferase reporter gene is a highly versatile tool for   in vitro  and   in vivo  assays and the corresponding reporter assay is extremely sensitive, reproducible. Therefore, luciferases have been employed as efficient reporters in a wide range of applications, including gene regulation and signaling, protein-protein interactions, drug screening, molecular imaging, cell-based assays, and noninvasive   in vivo  imaging [2-3].   Luciferases Luciferases originates from luminescent organisms, such as terrestrial and marine organisms. And here luciferases are mainly classified into two categories: D-luciferin-dependent luciferases and coelenterazine-dependent luciferases. D-luciferin-dependent luciferases Firefly