stiankog

• Kogler, Stian; Skottvoll, Frøydis Sved; Hruskova, Helena; Rise, Frode; Aizenshtadt, Aleksandra & Krauss, Stefan Johannes Karl [Show all 9 contributors for this article] (2025). Electromembrane Extraction Provides Unprecedented Selectivity for Drugs in Cell Culture Media Used in Organoid and Organ-on-Chip Systems. Analytical Chemistry. ISSN 0003-2700. 97(9), p. 4923–4931. doi: 10.1021/acs.analchem.4c04994. Full text in Research Archive Show summary

  The use of organoids and organ-on-chip technologies as nonanimal methodologies in drug discovery and personalized medicine is rapidly expanding. However, the complexity and small volumes of organoid culture samples present significant analytical challenges, e.g., in drug analysis using liquid chromatography–mass spectrometry (LC–MS). Essentially an electrophoresis across an oil membrane, electromembrane extraction (EME) offers a promising approach for measuring drugs, as it is, for example, compatible with small samples such as organoid and organ-on-chip formats. Given the potential of the technology, there is a need to assess the extraction purity of EME extracts to ensure EME’s compatibility with high-throughput, downstream analysis. This study evaluates the effectiveness of EME for sample cleanup in various common cell culture media used for organoids and organs-on-chips. The media were spiked with 90 small-molecule drugs. Using gel electrophoresis (sodium dodecyl sulfate polyacrylamide gel electrophoresis), high-resolution nuclear magnetic resonance spectroscopy, and LC–MS, we demonstrate that EME provides exhaustive removal of unwanted medium components (proteins, polar molecules, and apolar/neutral molecules) while selectively extracting the spiked small-molecule drugs. The approach was demonstrated with human stem-cell-derived liver organoids, allowing simple detection and monitoring of telltale cytochrome P450 metabolism. Taken together, our observations highlight an unprecedented ability of EME to provide sample cleanup for drug analysis in matrixes compatible with organoids and organ-on-chip technology.

• Kogler, Stian; Pedersen, Gustav Mathingsdal; Martínez-Ramírez, Felipe; Aizenshtadt, Aleksandra; Busek, Mathias & Krauss, Stefan Johannes Karl [Show all 8 contributors for this article] (2024). An FDA-Validated, Self-Cleaning Liquid Chromatography–Mass Spectrometry System for Determining Small-Molecule Drugs and Metabolites in Organoid/Organ-on-Chip Medium. Analytical Chemistry. ISSN 0003-2700. 96(29), p. 12129–12138. doi: 10.1021/acs.analchem.4c02246. Full text in Research Archive Show summary

  As organoids and organ-on-chip (OoC) systems move toward preclinical and clinical applications, there is an increased need for method validation. Using a liquid chromatography–mass spectrometry (LC–MS)-based approach, we developed a method for measuring small-molecule drugs and metabolites in the cell medium directly sampled from liver organoids/OoC systems. The LC–MS setup was coupled to an automatic filtration and filter flush system with online solid-phase extraction (SPE), allowing for robust and automated sample cleanup/analysis. For the matrix, rich in, e.g., protein, salts, and amino acids, no preinjection sample preparation steps (protein precipitation, SPE, etc.) were necessary. The approach was demonstrated with tolbutamide and its liver metabolite, 4-hydroxytolbutamide (4HT). The method was validated for analysis of cell media of human stem cell-derived liver organoids cultured in static conditions and on a microfluidic platform according to Food and Drug Administration (FDA) guidelines with regards to selectivity, matrix effects, accuracy, precision, etc. The system allows for hundreds of injections without replacing chromatography hardware. In summary, drug/metabolite analysis of organoids/OoCs can be performed robustly with minimal sample preparation.

• Kogler, Stian; Kømurcu, Kristina Sæterdal; Olsen, Christine Sandsnes; Shoji, Jun-ya; Skottvoll, Frøydis Sved & Krauss, Stefan Johannes Karl [Show all 8 contributors for this article] (2023). Organoids, organ-on-a-chip, separation science and mass spectrometry: An update. TrAC. Trends in analytical chemistry. ISSN 0165-9936. 161. doi: 10.1016/j.trac.2023.116996. Full text in Research Archive Show summary

  Organoids are 3D models of organs, grown in the laboratory from stem cells. An organ model grown/placed in microfluidic devices is commonly termed an “organ-on-a-chip”. Organoids and organ-on-a-chip devices are becoming important tools for studying physiology, disease modeling, drug discovery, personalized medicine, toxicology, and organ development/embryogenesis. We review how mass spectrometry is used for studying organoids and organ-on-a-chip-derived material. We first focus on proteomics, metabolomics/lipidomics, and hormones, typically discussing liquid chromatography-mass spectrometry (LC-MS) approaches. We then review work on mass spectrometry imaging (MSI) of organoids and discuss organ-on-a-chip coupled with mass spectrometry. The review focuses on research developments from the past four years. Mass spectrometric analysis of organoids and organ-on-a-chip has allowed novel insights on development and disease of e.g. brain, liver, and tumors, demonstrating potential for replacing or complementing animal models and other traditional model systems. Additional applications are emerging, e.g. related to sports doping and environmental toxicology.

• Kogler, Stian; Aizenshtadt, Aleksandra; Harrison, Sean; Skottvoll, Frøydis Sved; Berg, Henriette Engen & Abadpour, Shadab [Show all 14 contributors for this article] (2022). "Organ-in-a-Column" Coupled On-line with Liquid Chromatography-Mass Spectrometry. Analytical Chemistry. ISSN 0003-2700. 94(50), p. 17677–17684. doi: 10.1021/acs.analchem.2c04530. Full text in Research Archive Show summary

  Organoids, i.e., laboratory-grown organ models developed from stem cells, are emerging tools for studying organ physiology, disease modeling, and drug development. On-line analysis of organoids with mass spectrometry would provide analytical versatility and automation. To achieve these features with robust hardware, we have loaded liquid chromatography column housings with induced pluripotent stem cell (iPSC) derived liver organoids and coupled the “organ-in-a-column” units on-line with liquid chromatography-mass spectrometry (LC-MS). Liver organoids were coloaded with glass beads to achieve an even distribution of organoids throughout the column while preventing clogging. The liver organoids were interrogated “on column” with heroin, followed by on-line monitoring of the drug’s phase 1 metabolism. Enzymatic metabolism of heroin produced in the “organ-in-a-column” units was detected and monitored using a triple quadrupole MS instrument, serving as a proof-of-concept for on-line coupling of liver organoids and mass spectrometry. Taken together, the technology allows direct integration of liver organoids with LC-MS, allowing selective and automated tracking of drug metabolism over time.

• Kogler, Stian; Harrison, Sean; Aizenshtadt, Aleksandra; Skottvoll, Frøydis Sved; Abadpour, Shadab & Krauss, Stefan [Show all 10 contributors for this article] (2020). “Organoid-in-a-column” coupled on-line with liquid chromatography-mass spectrometry. BioRxiv. ISSN 0362-4331. doi: 10.1101/2020.09.08.282756. Full text in Research Archive

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• Kogler, Stian; Aizenshtadt, Aleksandra; Harrison, Sean Philip; Sullivan, Gareth John; Lundanes, Elsa & Bogen, Inger Lise [Show all 9 contributors for this article] (2023). Organ-in-a-column – automated on-line incubation, sampling, and LC-MS analysis for measurement of drug metabolites. Full text in Research Archive
• Røberg-Larsen, Hanne; Kømurcu, Kristina Sæterdal; Kogler, Stian; Olsen, Christine; Skottvoll, Frøydis Sved & Aizenshtadt, Aleksandra [Show all 8 contributors for this article] (2023). Organoids, Organ-on-a-chip and Mass Spectrometry. Full text in Research Archive
• Midtøy, Lise; Kømurcu, Kristina Sæterdal; Kogler, Stian; wilhelmsen, ingrid; Røberg-Larsen, Hanne & Wilson, Steven Ray Haakon (2023). Sensitive targeted proteomic analysis of cholesterol metabolism enzymes in hepatic stellate cells - going towards single cell analysis. Full text in Research Archive
• kvalvik, Eva; Kogler, Stian; Kømurcu, Kristina Sæterdal; Wilson, Steven Ray Haakon & Røberg-Larsen, Hanne (2023). Screening for biomarkers for non-alcoholic fatty liver disease using organ-in-a-column technology. Full text in Research Archive
• Haavaag, Jon Erlend; Kogler, Stian; Zawadzka, Malgorzata Elzbieta; Krauss, Stefan Johannes Karl; Wilson, Steven Ray Haakon & Røberg-Larsen, Hanne (2023). Building a high-resolution nano DESI-MS for organoid imaging: Analysis using a built and optimized imaging system. Full text in Research Archive
• Pedersen, Gustav Mathingsdal; Kogler, Stian & Røberg-Larsen, Hanne (2023). On-line Sample Preparation for "Organ-in-a-Column". Full text in Research Archive
• Tuner, Marius Bull; Kogler, Stian; Hansen, Frederik Andre; Pedersen-Bjergaard, Stig; Røberg-Larsen, Hanne & Wilson, Steven Ray Haakon (2023). Exploring electromembrane extraction and LC-MS for studying liver organoid metabolomics. Full text in Research Archive
• midtøy, Lise; Kømurcu, Kristina Sæterdal; Kogler, Stian; wilhelmsen, Ingrid; Røberg-Larsen, Hanne & Wilson, Steven Ray Haakon (2022). Sensitive targeted proteomic analysis of cholesterol metabolism enzymes in hepatic stellate cells - going towards single cell analysis. Full text in Research Archive
• Tuner, Marius Bull; Kogler, Stian; Hansen, Frederik Andre; Pedersen-Bjergaard, Stig; Røberg-Larsen, Hanne & Wilson, Steven Ray Haakon (2022). Exploring electromembrane extraction for sampling the organoid metabolome. Full text in Research Archive
• Kogler, Stian; Wilson, Steven Ray Haakon; Røberg-Larsen, Hanne; Krauss, Stefan Johannes Karl; Lundanes, Elsa & Skottvoll, Frøydis Sved (2021). Organ-in-a-column. Full text in Research Archive
• Kogler, Stian (2020). Organ on a chip - Kunstige organer, persontilpasset medisin og fremtidens forsøksdyr. Full text in Research Archive
• Kogler, Stian (2018). Tromsøstudent med på banebrytende organforskning. Full text in Research Archive
• Kogler, Stian & Steindal, Calin Constantin (2018). Identification of degradation markers in archaeological wood. Full text in Research Archive
• Pedersen, Gustav Mathingsdal; Røberg-Larsen, Hanne & Kogler, Stian (2023). Development of On-line Sample Clean-up System for Organ-in-a-Column. Universitetet i Oslo. Full text in Research Archive
• Kogler, Stian & Wilson, Steven Ray Haakon (2019). On-line “Organ-in-a-Column” Liquid Chromatography Electrospray Ionization Mass Spectrometry for Drug Metabolism Studies. Universitetet i Oslo. Full text in Research Archive Show summary

  Organoids and organ-on-a-chip (OoC) are human-induced pluripotent stem cell-derived miniaturized organs that recapitulate organ specific functions. They open up a plethora of opportunities to change the way we test drugs, research diseases, and treat patients. In this context, liquid chromatography (LC) and mass spectrometry (MS) will have to be used, e.g. for monitoring drug metabolism, toxicity screens and organ function. On-line coupling of these technologies would allow for automatized, high-throughput systems. By downsizing the LC-MS part, low abundant analytes can also be determined. In this study, an on-line sampling and analysis system for OoC-technology has been developed. The system features hepatic organoids in a flow-through tube with continuous medium flow (organ-in-a-column, OiC), a two-loop valve system for sampling, reverse phase liquid chromatography (LC) analysis, and electrospray ionization mass spectrometry (ESI-MS). Phase 1 metabolism of heroin to its metabolite morphine was successfully measured on-line with the described system. Live/dead double staining revealed the potential of long-term cultivation of organoids in the system. In parallel, tin oxide coated fused silica nanoelectrospray ionization (nanoESI) emitters suitable for the most narrow bore chromatographic columns have been developed with HF etching and an atomic layer deposition (ALD) approach. The emitters have been tested against commercially available stainless steel (SS) emitters by injections of heroin and showed similar signal-to-noise ratios. These emitters will find a use in a future miniaturized version of the system described herein. In summary, a proof-of-concept system for on-line measurement of heroin metabolites has successfully been developed for a simple OoC-like device. NanoESI emitters for the most narrow columns have also been developed with a conductive ALD-coating of SnO2.

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