TY  - JOUR
A1  - Votteler, Miriam
A1  - Carvajal Berrio, Daniel A.
A1  - Pudlas, Marieke
A1  - Walles, Heike
A1  - Schenke-Layland, Katja
T1  - Non-contact, Label-free Monitoring of Cells and Extracellular Matrix using Raman Spectroscopy
JF  - Journal of Visual Expression
N2  - Non-destructive, non-contact and label-free technologies to monitor cell and tissue cultures are needed in the field of biomedical research.1-5 However, currently available routine methods require processing steps and alter sample integrity. Raman spectroscopy is a fast method that enables the measurement of biological samples without the need for further processing steps. This laser-based technology detects the inelastic scattering of monochromatic light.6 As every chemical vibration is assigned to a specific Raman band (wavenumber in cm-1), each biological sample features a typical spectral pattern due to their inherent biochemical composition.7-9 Within Raman spectra, the peak intensities correlate with the amount of the present molecular bonds.1 Similarities and differences of the spectral data sets can be detected by employing a multivariate analysis (e.g. principal component analysis (PCA)).10

Here, we perform Raman spectroscopy of living cells and native tissues. Cells are either seeded on glass bottom dishes or kept in suspension under normal cell culture conditions (37 °C, 5% CO2) before measurement. Native tissues are dissected and stored in phosphate buffered saline (PBS) at 4 °C prior measurements. Depending on our experimental set up, we then either focused on the cell nucleus or extracellular matrix (ECM) proteins such as elastin and collagen. For all studies, a minimum of 30 cells or 30 random points of interest within the ECM are measured. Data processing steps included background subtraction and normalization.
KW  - tissue engineering
KW  - label-free analysis
KW  - raman spectroscopy
KW  - bioengineering
KW  - living cells
KW  - extracellular matrix
Y1  - 2012
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-124569
VL  - 63
IS  - e3977
ER  - 
TY  - JOUR
A1  - Fecher, David
A1  - Hofmann, Elisabeth
A1  - Buck, Andreas
A1  - Bundschuh, Ralph
A1  - Nietzer, Sarah
A1  - Dandekar, Gudrun
A1  - Walles, Thorsten
A1  - Walles, Heike
A1  - Lückerath, Katharina
A1  - Steinke, Maria
T1  - Human Organotypic Lung Tumor Models: Suitable For Preclinical \(^{18}\)F-FDG PET-Imaging
JF  - PLoS ONE
N2  - Development of predictable in vitro tumor models is a challenging task due to the enormous complexity of tumors in vivo. The closer the resemblance of these models to human tumor characteristics, the more suitable they are for drug-development and –testing. In the present study, we generated a complex 3D lung tumor test system based on acellular rat lungs. A decellularization protocol was established preserving the architecture, important ECM components and the basement membrane of the lung. Human lung tumor cells cultured on the scaffold formed cluster and exhibited an up-regulation of the carcinoma-associated marker mucin1 as well as a reduced proliferation rate compared to respective 2D culture. Additionally, employing functional imaging with 2-deoxy-2-[\(^{18}\)F]fluoro-D-glucose positron emission tomography (FDG-PET) these tumor cell cluster could be detected and tracked over time. This approach allowed monitoring of a targeted tyrosine kinase inhibitor treatment in the in vitro lung tumor model non-destructively. Surprisingly, FDG-PET assessment of single tumor cell cluster on the same scaffold exhibited differences in their response to therapy, indicating heterogeneity in the lung tumor model. In conclusion, our complex lung tumor test system features important characteristics of tumors and its microenvironment and allows monitoring of tumor growth and -metabolism in combination with functional imaging. In longitudinal studies, new therapeutic approaches and their long-term effects can be evaluated to adapt treatment regimes in future.
KW  - lung and intrathoracic tumors
KW  - trachea
KW  - adenocarcinoma of the lung
KW  - cancer treatment
KW  - secondary lung tumors
KW  - pulmonary imaging
KW  - extracellular matrix
KW  - collagens
Y1  - 2016
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-179678
VL  - 11
IS  - 8
ER  -