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ePaper created 2016-03-17, 15:06:18 | version 1.45.01
68 CARDIAC STEM CELL DIFFERENTIATION AND NON-INVASIVE MONITORING Eva Brauchle, Nian Shen, Shannon Layland, Svenja Hinderer, Katja Schenke-Layland Challenges in cardiovascular disease Despite a multitude of newly developed drugs and diag- nostics, and significant advances in cardiology and cardiac surgery, diseases of the cardiovascular system are still the number one killer worldwide according to figures from the World Health Organization. Here, heart muscle or heart valves are affected frequently. Damage to these tissues may have different causes; for example, risk factors such as smoking and unhealthy diet can lead to a heart attack. Congenital diseases are also known to be the cause of organ or tissue failure. In adult humans, the heart does not regenerate after damage, which significantly reduces the performance of the heart, and thus the mortality and quality of life of these patients. A num- ber of scientists are working towards restoring heart function through regeneration or developing cardiac test systems, which would enable the in vitro testing of new therapies and drugs in a humanized model. Primary isolated beating heart muscle cells cannot be cultured in vitro. Therefore, stem cell derived heart muscle cells are required to create a functional heart muscle test system. Human heart muscle test system from embryonic stem cells cientific studies have demonstrated that embryonic stem cells differentiate into functional cardiomyocytes under spe- cific chemical culture conditions Our strategy is to create a physiological environment that will differentiate and mature cardiomyocytes e have developed a stretch-flow bioreactor which enables the dynamic culture of embryonic stem cells under defined tensile and continuous medium flow e have demonstrated that we can differentiate and mature beating cardiomyocytes capable of calcium transport with biome- chanical cues from the flow and stretch The addition of caf- feine into the dynamic culture increaes cardiomyocyte stroke rate and influ of calcium ions The calcium channel blocker Verapamil has the opposite effect when added to the culture. In current studies, a carrier substrate is being used to establish a three-dimensional culture similar to our previous work us- ing electrospun substrates for 3D culture of smooth muscle cells [1] and heart valve cells [2] with the aim of creating a 3D myocard test system. Marker-free monitoring of cardiac stem cell differentiation The identification of cell differentiating from embryonic stem cells to myocardial cells is an important step in the prepara- tion of a test system. Classically, protein expression is used to identify different stages of embryonic stem cell differentiation. However, this alters the cells making them unsuitable for con- tinuous monitoring of the differentiation. Raman microspectroscopy Raman microspectroscopy is an optical technology that can be used for marker-free global analysis of biological cell and tissue samples. It is based on the effect of scattered light, wherein few photons of the incident monochromatic light are shifted by molecular vibration within a sample. Raman spectroscopy detects a molecular fingerprint of an ob ect As an object changes, its Raman spectra typically changes as well. Thus, cell differentiation processes can be non-invasively monitored and the uni ue profiles of different cell types and states from a stem cell, or any cell, can be distinguished. 1 MEDICINE