Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2023 Nov;44(21-22):1682-1697.
doi: 10.1002/elps.202300095. Epub 2023 Aug 13.

Simultaneous LC-MS determination of glucose regulatory peptides secreted by stem cell-derived islet organoids

Christine Olsen  1   2 , Chencheng Wang  2   3 , Aleksandra Aizenshtadt  2 , Shadab Abadpour  2   3 , Elsa Lundanes  1 , Frøydis Sved Skottvoll  4 , Alexey Golovin  5 , Mathias Busek  2 , Stefan Krauss  2   5 , Hanne Scholz  2   3 , Steven Ray Wilson  2   3
Affiliations

Simultaneous LC-MS determination of glucose regulatory peptides secreted by stem cell-derived islet organoids

Christine Olsen et al. Electrophoresis. 2023 Nov.

Abstract

For studying stem cell-derived islet organoids (SC-islets) in an organ-on-chip (OoC) platform, we have developed a reversed-phase liquid chromatography-tandem mass spectrometry (RPLC-MS/MS) method allowing for simultaneous determination of insulin, somatostatin-14, and glucagon, with improved matrix robustness compared to earlier methodology. Combining phenyl/hexyl-C18 separations using 2.1 mm inner diameter LC columns and triple quadrupole mass spectrometry, identification and quantification were secured with negligible variance in retention time and quantifier/qualifier ratios, negligible levels of carryover (<2%), and sufficient precision (±10% RSD) and accuracy (±15% relative error) with and without use of an internal standard. The obtained lower limits of quantification were 0.2 µg/L for human insulin, 0.1 µg/L for somatostatin-14, and 0.05 µg/L for glucagon. The here-developed RPLC-MS/MS method showed that the SC-islets have an insulin response dependent on glucose concentration, and the SC-islets produce and release somatostatin-14 and glucagon. The RPLC-MS/MS method for these peptide hormones was compatible with an unfiltered offline sample collection from SC-islets cultivated on a pumpless, recirculating OoC (rOoC) platform. The SC-islets background secretion of insulin was not significantly different on the rOoC device compared to a standard cell culture well-plate. Taken together, RPLC-MS/MS method is well suited for multi-hormone measurements of SC-islets on an OoC platform.

Keywords: glucose regulatory hormones; liquid chromatography-tandem mass spectrometry; organ-on-chip; stem cell-derived islet organoids.

PubMed Disclaimer

References

REFERENCES

    1. Tang XY, Wu S, Wang D, Chu C, Hong Y, Tao M, et al. Human organoids in basic research and clinical applications. Signal Transduct Target Ther. 2022;7(1):168.
    1. Kim J, Koo BK, Knoblich JA. Human organoids: Model systems for human biology and medicine. Nat Rev Mol Cell Biol. 2020;21(10):571-84.
    1. Ingber DE. Human organs-on-chips for disease modelling, drug development and personalized medicine. Nat Rev Genet. 2022;23(8):467-91.
    1. Busek M, Aizenshtadt A, Koch T, Frank A, Delon L, Martinez MA, et al. Pump-less, recirculating organ-on-a-chip (rOoC) platform. Lab Chip. 2023;23:591-608.
    1. Lin A, Sved Skottvoll F, Rayner S, Pedersen-Bjergaard S, Sullivan G, Krauss S, et al. 3D cell culture models and organ-on-a-chip: meet separation science and mass spectrometry. Electrophoresis. 2020;41(1-2):56-64.

Publication types

LinkOut - more resources