A simple and sensitive assay for GH activity based on 3T3-F442A cell differentiation.
We describe a fast, sensitive, specific, and simple in vitro assay for GH biological activity, based on the differentiation of 3T3-F442A cells into adipocytes. The 3T3-F442A cells were directly plated at 1.5 x 10(4)cells/cm(2) in medium with or without various concentrations of human growth hormone (hGH). After 7 days, cells were lysed with buffer containing 0.5 % (v/v) Triton X-100, and adipose conversion was quantitated by the activity of the adipogenic enzyme glycerophosphate dehydrogenase. The assay is highly sensitive and specific for GH from different species. These culture conditions have shortened the time for the cells to undergo adipose differentiation, and they might also be useful to design and test drugs or agents that modify adipocyte differentiation or lipid metabolism, or for evaluation of cytotoxic and pharmacologic effects of drugs and other compounds.
Juárez-Aguilar
Enrique
E
Department of Cell Biology, Centro de Investigación y de Estudios Avanzados del I.P.N., Apdo. Postal 14-740, 07000, México, D.F., Mexico.
Castro-Muñozledo
Federico
F
Kuri-Harcuch
Walid
W
D016475:3T3 Cells; D017667:Adipocytes / Q000166:cytology / Q000187:drug effects* / Q000201:enzymology* / Q000502:physiology; D000818:Animals; D015374:Biosensing Techniques; D002454:Cell Differentiation / Q000187:drug effects*; D004305:Dose-Response Relationship, Drug; D004789:Enzyme Activation; D005993:Glycerolphosphate Dehydrogenase / Q000032:analysis / Q000378:metabolism*; D013006:Growth Hormone / Q000032:analysis* / Q000494:pharmacology*; D051379:Mice; D012098:Reproduction; D012680:Sensitivity and Specificity
D013006:Growth Hormone; D005993:Glycerolphosphate Dehydrogenase