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  - Hinderer, Svenja
A1  - Shen, Nian
A1  - Ringuette, Léa-Jeanne
A1  - Hansmann, Jan
A1  - Reinhardt, Dieter P
A1  - Brucker, Sara Y
A1  - Davis, Elaine C
A1  - Schenke-Layland, Katja
T1  - In vitro elastogenesis: instructing human vascular smooth muscle cells to generate an elastic fiber-containing extracellular matrix scaffold
JF  - Biomedical Materials
N2  - Elastic fibers are essential for the proper function of organs including cardiovascular tissues such as heart valves and blood vessels. Although (tropo)elastin production in a tissue-engineered construct has previously been described, the assembly to functional elastic fibers in vitro using human cells has been highly challenging. In the present study, we seeded primary isolated human vascular smooth muscle cells (VSMCs) onto 3D electrospun scaffolds and exposed them to defined laminar shear stress using a customized bioreactor system. Increased elastin expression followed by elastin deposition onto the electrospun scaffolds, as well as on newly formed fibers, was observed after six days. Most interestingly, we identified the successful deposition of elastogenesis-associated proteins, including fibrillin-1 and -2, fibulin-4 and -5, fibronectin, elastin microfibril interface located protein 1 (EMILIN-1) and lysyl oxidase (LOX) within our engineered constructs. Ultrastructural analyses revealed a developing extracellular matrix (ECM) similar to native human fetal tissue, which is composed of collagens, microfibrils and elastin. To conclude, the combination of a novel dynamic flow bioreactor and an electrospun hybrid polymer scaffold allowed the production and assembly of an elastic fiber-containing ECM.
KW  - elastin
KW  - elastic fibers
KW  - electrospinning
KW  - tissue engineering
KW  - regenerative medicine
KW  - heart valve
KW  - cardiovascular
Y1  - 2015
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-254074
VL  - 10
IS  - 3
ER  -