Xf24 respirometry machine

Manufactured by Agilent Technologies

The XF24 respirometry machine is a laboratory equipment designed to measure the oxygen consumption and extracellular acidification rates of cells. It provides real-time data on cellular metabolism and bioenergetics.

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Lab products found in correlation

Seahorse xf assay medium, by Agilent Technologies (2 mentions)

2 protocols using xf24 respirometry machine

Kidney Organoid Respiration Profiling

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Kidney organoids at day 16 of differentiation were resuspended in warm Seahorse XF Assay Medium (Seahorse Bioscience). Individual organoids were transferred to an islet plate (one organoid per well) containing 400 μl of medium per well. After 1 h of incubation at 37 °C, plates were loaded into an XF24 respirometry machine (Seahorse Bioscience). Uncoupled and maximum OCR were assayed with oligomycin (1 μM) and FCCP (1.5 μM). To inhibit complex I- and III-dependent respiration, rotenone (1 μM) and antimycin A (1 μM) were used, respectively. OCR represents the oxygen tension and acidification of the medium as a function of time (pmol min⁻¹).

Garreta E., Prado P., Tarantino C., Oria R., Fanlo L., Martí E., Zalvidea D., Trepat X., Roca-Cusachs P., Gavaldà-Navarro A., Cozzuto L., Campistol J.M., Belmonte J.C., del Pozo C.H, & Montserrat N. (2019). Fine tuning the extracellular environment accelerates the derivation of kidney organoids from human pluripotent stem cells. Nature materials, 18(4), 397-405.

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Measuring OCR in Kidney Cell Types

Cited 2 times

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The measurement of OCR in kidney organoid-derived cells and proximal tubular cells was performed using an XFe24 extracellular flux analyzer (Seahorse Bioscience) as previously described (Dhillon et al., 2021 (link); Garreta et al., 2019 (link)). Cells were plated at density of 20,000 cells/well in a Seahorse cell culture microplate in warm Seahorse XF Assay Medium supplemented with glucose 5mM, pyruvate 0.5mM and glutamine 2mM (Seahorse Bioscience). After 1 h of incubation at 37 °C, plates were loaded into an XF24 respirometry machine (Seahorse Bioscience). Uncoupled and maximum OCR were assayed with oligomycin (10 μM) and FCCP (2 μM). To inhibit complex I- and III-dependent respiration, rotenone (5 μM) and antimycin A (15 μM) were used, respectively. OCR represents the oxygen tension and acidification of the medium as a function of time (pmol min−1). OCR was normalized to protein quantity in each well.

Garreta E., Prado P., Stanifer M.L., Monteil V., Marco A., Ullate-Agote A., Moya-Rull D., Vilas-Zornoza A., Tarantino C., Romero J.P., Jonsson G., Oria R., Leopoldi A., Hagelkruys A., Gallo M., González F., Domingo-Pedrol P., Gavaldà A., del Pozo C.H., Hasan Ali O., Ventura-Aguiar P., Campistol J.M., Prosper F., Mirazimi A., Boulant S., Penninger J.M, & Montserrat N. (2022). A diabetic milieu increases ACE2 expression and cellular susceptibility to SARS-CoV-2 infections in human kidney organoids and patient cells. Cell Metabolism, 34(6), 857-873.e9.

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