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A list of all pages that have property "Has abstract" with value "Quality control is required in clinical studies to ensure accuracy and reproducibility. The technical repeatability of subsamples reflects instrumental resolution and experimental precision, but is insufficiently reported in mitochondrial respiratory research. A frequently neglected problem is quality control of chemicals and incubation media. Mitochondrial respiration medium MiR05 is prepared from MiR05-Kit (Oroboros Instruments), which contains 7 chemicals and is stored as a crystalline powder. The powder is dissolved in H2O, bovine serum albumin (BSA) is added, and the pH is adjusted. We evaluated the quality of 5 lots of MiR05-Kit stored at room temperature for 2 months to 3 years. Two substrate-uncoupler-inhibitor titration (SUIT) reference protocols [1] were applied in the Oroboros O2k with cryopreserved HEK293T cells. Compared to the instrumental limit of detection of volume-specific O2 flux of ±1 pmol·s-1·mL-1, the range of technical repeats in ROUTINE respiration was ±5 pmol·s-1·mL-1 or ±15 % using 8-16 O2k chambers in parallel at a cell concentration of 106 x∙mL-1. The biological variability between cell batches was higher. Homogenous stocks of suspended cells were used to compare MiR05 lots at 3 annual intervals. The MiR05-Kit was stable up to 3 years as shown by consistent cell respiration with different MiR05 lots. Since many different pathway and coupling control states of living and permeabilized cells were covered in the 2 SUIT protocols [1], these results exclude specific inhibitory modifications of MiR05 during storage. Our study demonstrates a constant quality of MiR05-Kit up to 3 years of storage. This represents an important quality control in high-resolution respirometry to obtain reliable and reproducible results in scientific and clinical investigations.". Since there have been only a few results, also nearby values are displayed.

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Baglivo 2022a ESCI Bari + (Quality control is required in clinical st … Quality control is required in clinical studies to ensure accuracy and reproducibility. The technical repeatability of subsamples reflects instrumental resolution and experimental precision, but is insufficiently reported in mitochondrial respiratory research. A frequently neglected problem is quality control of chemicals and incubation media. Mitochondrial respiration medium MiR05 is prepared from MiR05-Kit (Oroboros Instruments), which contains 7 chemicals and is stored as a crystalline powder. The powder is dissolved in H2O, bovine serum albumin (BSA) is added, and the pH is adjusted. We evaluated the quality of 5 lots of MiR05-Kit stored at room temperature for 2 months to 3 years. Two substrate-uncoupler-inhibitor titration (SUIT) reference protocols [1] were applied in the Oroboros O2k with cryopreserved HEK293T cells. Compared to the instrumental limit of detection of volume-specific O2 flux of ±1 pmol·s-1·mL-1, the range of technical repeats in ROUTINE respiration was ±5 pmol·s-1·mL-1 or ±15 % using 8-16 O2k chambers in parallel at a cell concentration of 106 x∙mL-1. The biological variability between cell batches was higher. Homogenous stocks of suspended cells were used to compare MiR05 lots at 3 annual intervals.The MiR05-Kit was stable up to 3 years as shown by consistent cell respiration with different MiR05 lots. Since many different pathway and coupling control states of living and permeabilized cells were covered in the 2 SUIT protocols [1], these results exclude specific inhibitory modifications of MiR05 during storage.Our study demonstrates a constant quality of MiR05-Kit up to 3 years of storage. This represents an important quality control in high-resolution respirometry to obtain reliable and reproducible results in scientific and clinical investigations.ant quality control in high-resolution respirometry to obtain reliable and reproducible results in scientific and clinical investigations.)

Baglivo 2022a ESCI Bari +