TY  - JOUR
A1  - Dong, Meng
A1  - Böpple, Kathrin
A1  - Thiel, Julia
A1  - Winkler, Bernd
A1  - Liang, Chunguang
A1  - Schueler, Julia
A1  - Davies, Emma J.
A1  - Barry, Simon T.
A1  - Metsalu, Tauno
A1  - Mürdter, Thomas E.
A1  - Sauer, Georg
A1  - Ott, German
A1  - Schwab, Matthias
A1  - Aulitzky, Walter E.
T1  - Perfusion air culture of precision-cut tumor slices: an ex vivo system to evaluate individual drug response under controlled culture conditions
JF  - Cells
N2  - Precision-cut tumor slices (PCTS) maintain tissue heterogeneity concerning different cell types and preserve the tumor microenvironment (TME). Typically, PCTS are cultured statically on a filter support at an air–liquid interface, which gives rise to intra-slice gradients during culture. To overcome this problem, we developed a perfusion air culture (PAC) system that can provide a continuous and controlled oxygen medium, and drug supply. This makes it an adaptable ex vivo system for evaluating drug responses in a tissue-specific microenvironment. PCTS from mouse xenografts (MCF-7, H1437) and primary human ovarian tumors (primary OV) cultured in the PAC system maintained the morphology, proliferation, and TME for more than 7 days, and no intra-slice gradients were observed. Cultured PCTS were analyzed for DNA damage, apoptosis, and transcriptional biomarkers for the cellular stress response. For the primary OV slices, cisplatin treatment induced a diverse increase in the cleavage of caspase-3 and PD-L1 expression, indicating a heterogeneous response to drug treatment between patients. Immune cells were preserved throughout the culturing period, indicating that immune therapy can be analyzed. The novel PAC system is suitable for assessing individual drug responses and can thus be used as a preclinical model to predict in vivo therapy responses.
KW  - precision-cut tumor slices
KW  - perfusion culture
KW  - tumor microenvironment
KW  - ovarian tumor
KW  - individual drug responses
KW  - mouse xenografts
KW  - preclinical model
KW  - personalized medicine
Y1  - 2023
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-311030
SN  - 2073-4409
VL  - 12
IS  - 5
ER  - 
TY  - JOUR
A1  - Volland, Julian Manuel
A1  - Kaupp, Johannes
A1  - Schmitz, Werner
A1  - Wünsch, Anna Chiara
A1  - Balint, Julia
A1  - Möllmann, Marc
A1  - El-Mesery, Mohamed
A1  - Frackmann, Kyra
A1  - Peter, Leslie
A1  - Hartmann, Stefan
A1  - Kübler, Alexander Christian
A1  - Seher, Axel
T1  - Mass spectrometric metabolic fingerprinting of 2-Deoxy-D-Glucose (2-DG)-induced inhibition of glycolysis and comparative analysis of methionine restriction versus glucose restriction under perfusion culture in the murine L929 model system
JF  - International Journal of Molecular Sciences
N2  - All forms of restriction, from caloric to amino acid to glucose restriction, have been established in recent years as therapeutic options for various diseases, including cancer. However, usually there is no direct comparison between the different restriction forms. Additionally, many cell culture experiments take place under static conditions. In this work, we used a closed perfusion culture in murine L929 cells over a period of 7 days to compare methionine restriction (MetR) and glucose restriction (LowCarb) in the same system and analysed the metabolome by liquid chromatography mass spectrometry (LC-MS). In addition, we analysed the inhibition of glycolysis by 2-deoxy-D-glucose (2-DG) over a period of 72 h. 2-DG induced very fast a low-energy situation by a reduced glycolysis metabolite flow rate resulting in pyruvate, lactate, and ATP depletion. Under perfusion culture, both MetR and LowCarb were established on the metabolic level. Interestingly, over the period of 7 days, the metabolome of MetR and LowCarb showed more similarities than differences. This leads to the conclusion that the conditioned medium, in addition to the different restriction forms, substantially reprogramm the cells on the metabolic level.
KW  - amino acid restriction
KW  - glucose restriction
KW  - mass spectrometry
KW  - low carb
KW  - 2-deoxy-D-glucose
KW  - 2-DG
KW  - methionine
KW  - perfusion culture
KW  - energy restriction
KW  - caloric restriction
Y1  - 2022
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-286007
SN  - 1422-0067
VL  - 23
IS  - 16
ER  -