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Evaluation of Preparation Techniques for Optimal Hepcidin Detection in Human Hepatocyte Culture
Author(s) -
Morehouse Zachary Patrick,
Easparro Brandon,
Proctor Caleb,
Nash Rodney J.,
Atwood James
Publication year - 2018
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.2018.32.1_supplement.873.11
Subject(s) - hepcidin , erythropoiesis , medicine , hereditary hemochromatosis , ferroportin , anemia , erythropoietin , hemochromatosis , kidney disease , physiology , endocrinology
Approximately 40% of adults aged 65 and older are affected by Chronic Kidney Disease (CKD). Anemia is an associated symptom seen in many patients diagnosed with CKD, promoting elevated cardiovascular risk, decreased quality of life, and increased morbidity and mortality in affected populations. Hepcidin‐25 (hepcidin) is a key iron regulating hormone secreted by hepatocytes to regulate and maintain body iron homeostasis and dietary iron absorption. Hepcidin does this by stifling iron release from macrophages, preventing the normal recycling of iron in the body needed to support erythropoiesis. Elevated levels of hepcidin have been observed in both adults and children suffering from CKD, while its pathological mechanism of action is still relatively unstudied. There is a growing need to further the understanding of hepcidin's role in CDK and other iron related pathologies. Currently there is no standardized methodology for hepcidin evaluation studies on relevant human cell lines, and there is a growing need for studies to be done in this field to assist biomedical researchers in the translation of clinically relevant lab findings from the benchtop to the bedside. Hep G2 is a perpetual human hepatic cancer cell line confirmed to produce hepcidin and a commonly used lineage when studying hepatic function in vitro . While it is confirmed that Hep G2 cells produce hepcidin when cultured, there is currently no standardized procedure for detection of this peptide in vitro from these, or any other human derived cell lines. The expanding clinical and biomedical implications of hepcidin has prompted the need to determine an optimal method for detection of hepcidin in vitro . In this study we use of Hep G2 cells as a model for hepcidin peptide detection and various tissue processing techniques to prepare the cells hepcidin quantification via ELISA. This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal .