Measurement of urea synthesis in cultured stem cell-derived hepatocyte-like cells
Scope of the method
- Human health
- Basic Research
- In vitro - Ex vivo
- Human derived cells / tissues / organs
- Stem cells
- hepatic toxicity
- Cell culture
- cellular programming
The present standard procedure describes a protocol for measuring the urea concentration in supernatant of human stem cell-derived hepatocyte-like cells. This procedure relies on a chromogenic reagent that forms a colored complex specifically with urea. The latter can be measured and is directly proportional to the urea concentration in the sample.
- Biosafety cabinet ;
- Multiplate reader ;
- Thermostated bath.
- History of use
- Internally validated
Pros, cons & Future potential
The current protocol represents a simple and direct method to quantitatively measure the urea concentration in human stem cell-derived hepatocyte-like cell cultures. This assay has no harmful effect on the cultured cells. Therefore, after incubation of the cells with the substrate (ammonium chloride (NH4Cl)), the cultures can be maintained.
Can be applied to other types of in vitro systems of hepatocytes.
References, associated documents and other information
Gómez-Lechón MJ, Lahoz A, Gombau L, Castell JV, Donato MT. (2010) In vitro evaluation of potential hepatotoxicity induced by drugs. Curr Pharm Des 16:1963-77
Lavon N, Benvenisty N. (2005) Study of hepatocyte differentiation using embryonic stem cells. J Cell Biochem 96:1193-202
Roelandt P, Pauwelyn KA, Sancho-Bru P, Subramanian K, Ordovas L, Vanuytsel K, Geraerts M, Firpo M, De Vos R, Fevery J, Nevens F, Hu WS, Verfaillie CM. (2010) Human Embryonic and Rat Adult Stem Cells with Primitive Endoderm-Like Phenotype Can Be Fated to Definitive Endoderm, and Finally Hepatocyte-Like Cells. PLoS One; 5(8): e12101
Snykers S, Vanhaecke T, De Becker A, Papeleu P, Vinken M, Van Riet I, Rogiers V. (2007) Chromatin remodeling agent trichostatin A: a key-factor in the hepatic differentiation of human mesenchymal stem cells derived of adult bone marrow. BMC Dev Biol 7:24
Snykers S, De Kock J, Rogiers V, Vanhaecke T. (2009) In vitro differentiation of embryonic and adult stem cells into hepatocytes: state of the art. Stem Cells 27:577-605
OrganisationsVrije Universiteit Brussel
Pharmaceutical and Pharmacological Sciences (FARM)
In Vitro Toxicology and Dermato-cosmetology