ABSTRACTS â ORAL PRESENTATIONS - AMCA, spol. s r.o.
ABSTRACTS â ORAL PRESENTATIONS - AMCA, spol. s r.o. ABSTRACTS â ORAL PRESENTATIONS - AMCA, spol. s r.o.
P84. THE ROLE OF ATP-BINDING TRANSPORTERS ASSOCIATED WITH MULTI-DRUG RESISTANCE IN STEM CELLS Martina Lánová 1 , Josef Večeřa 1 , Jan Kučera 1 , Jiřina Procházková 1 , Jiří Pacherník 1,2 1 Institute of Experimental Biology, Faculty of Science, Masaryk University, Brno, Czech Republic 2 International Clinical Research Center – Centre of Biomolecular and Cellular Engineering, St. Anne’s University Hospital Brno, Czech Republic ATP-binding transporters (ABC-t) play various roles in regulation organism function and homeostasis from prokaryota to mammals. ABC-t mediate transport of mainly lipophilic substances through cellular membranes. Some ABC-t are important in cell protection against endogenous and importantly also exogenous toxins. These transporters are called ABC-t associated with multi-drug resistance (ABC-t/MDR), according to their role in resistance of tumor cells to pharmacotherapy. ABC-t/MDR are also over-expressed in stem cells, where their protective role is expected, too. Particularly, ABCB1, ABCC1, and ABCG2 are common ABC-t/MDR expressed in stem cells. However, substrates of ABC-t/ MDR are not only toxins, but also important signaling molecules as well leukotrienes and/or glutathione conjugates and porphyrins, which mediated balance in intracellular oxidation-reduction processing. Thus we hypothesize the role of ABC-t/MDR also in regulation of stem cells fate. To test this hypothesis we analyzed effect of modulation of ABC-t/MDR activity in embryonic and neural stem cells. We observed that ABCC1 and ABCG2 are the most expressed ABC-t/MDR in our tested stem cells. Importantly, inhibition of these ABC-t/MDR leads to decreasing of stemness and induction of differentiation in both embryonic and neural stem cells. Analysis of mechanism of observed effect and identification of studying ABC-t/MDR substrates, which may be responsible for this effect, are in progression. Acknowledgment This work was supported by grant from MEYS CR Cost CZ LD11015 and Ministry of Education, Youth and Sport of the Czech Republic MSM0021622430 P85. CYTOKINE SECRETION DURING NEURAL PROGENITOR CELL DIFFERENTIATION Karla Jarkovska 1 , Katerina Mairychova 1 , Rita Hrabakova 1 , Jirina Tyleckova 1 , Martin Marsala 2 , Silvia Marsala 2 and Hana Kovarova 1 1 Institute of Animal Physiology and Genetics, AS CR v.v.i., Libechov, Czech Republic 2 University of California, San Diego, United States of America Neural stem cells have an enormous potential in the therapy of the neurodegenerative diseases or spinal cord injuries. For therapeutic applications it is essential to characterise the developmental stages of neural cells from proliferating progenitors to fully differentiated neural cells. However, suitable biomarkers indicating commitment to a Analytical Cytometry VII 189
given cell type are missing and these may facilitate the selection of optimal clones for cell transplantation In addition, the demands on treatment of neurodegenerative diseases are expected to rise due to the increase in long-aged human population and higher disease incidence. During the last two decades, stem cells discovery revolutionised the search for new therapies and possible neural stem cells transplantation is a focus for many researchers, some of them reporting significant function improvement after transplantation (Lu et al., 2012). The aim of stem cells characterisation is, inter alia, to identify molecules monitoring post-transplantation stages in the course of cell-based therapies. The proteins secreted by cells, called secretome, offer such opportunities, hence the aim of this study is characterisation of secretome of neural progenitor cells and its changes during neural differentiation. The human neural stem cells derived from foetal spinal cord were isolated by FACS according to described cell surface signatures from heterogeneous population (Yuan et al., 2011). Sorted cells were cultivated in monolayer by bFGF and subsequently differentiated by adding BDNF and GDNF for 30 days. Media were collected every other day of neuronal differentiation. For the quantification of the cytokines level we selected the Luminex xMAP approach followed by statistical evaluation and interaction network modeling. The most significant quantitative changes were observed in the level of 10 cytokines (CCL2, CCL3, CCL5, CXCL1, CXCL5, CXCL8, IL-10, MCSF, VEGF, angiogenin) with three different cytokine secretion profiles during differentiation. Our findings suggest a set of new cytokines which could help in monitoring of precursor cells differentiation and survival as well as therapeutic effect. Despite that, further studies on stem cells secreted proteins are needed. Acknowledgements This study was supported by Technical Agency of the Czech Republic (TA01011466) and the project ExAM from European Regional Development Fund CZ.1.05/2.1.00/03.0124. References Lu, P., Wang, Y., Graham, L., McHale, K., Gao, M., Wu, D., Brock, J., Blesch, A., Rosenzweig, E.S., Havton, L.A., et al. (2012). Long-Distance Growth and Connectivity of Neural Stem Cells after Severe Spinal Cord Injury. Cell 150, 1264–1273. Yuan, S.H., Martin, J., Elia, J., Flippin, J., Paramban, R.I., Hefferan, M.P., Vidal, J.G., Mu, Y., Killian, R.L., Israel, M.A., et al. (2011). Cell-Surface Marker Signatures for the Isolation of Neural Stem Cells, Glia and Neurons Derived from Human Pluripotent Stem Cells. PLoS ONE 6, e17540. Corresponding author: Katerina Mairychova, katerina.mairychova@gmail.com 190 Analytical Cytometry VII
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P84. THE ROLE OF ATP-BINDING TRANSPORTERS ASSOCIATED WITH MULTI-DRUG<br />
RESISTANCE IN STEM CELLS<br />
Martina Lánová 1 , Josef Večeřa 1 , Jan Kučera 1 , Jiřina Procházková 1 , Jiří Pacherník 1,2<br />
1<br />
Institute of Experimental Biology, Faculty of Science, Masaryk University, Brno, Czech<br />
Republic<br />
2<br />
International Clinical Research Center – Centre of Biomolecular and Cellular<br />
Engineering, St. Anne’s University Hospital Brno, Czech Republic<br />
ATP-binding transporters (ABC-t) play various roles in regulation organism function and<br />
homeostasis from prokaryota to mammals. ABC-t mediate transport of mainly lipophilic<br />
substances through cellular membranes. Some ABC-t are important in cell protection<br />
against endogenous and importantly also exogenous toxins. These transporters are<br />
called ABC-t associated with multi-drug resistance (ABC-t/MDR), according to their role<br />
in resistance of tumor cells to pharmacotherapy. ABC-t/MDR are also over-expressed in<br />
stem cells, where their protective role is expected, too. Particularly, ABCB1, ABCC1, and<br />
ABCG2 are common ABC-t/MDR expressed in stem cells. However, substrates of ABC-t/<br />
MDR are not only toxins, but also important signaling molecules as well leukotrienes<br />
and/or glutathione conjugates and porphyrins, which mediated balance in intracellular<br />
oxidation-reduction processing. Thus we hypothesize the role of ABC-t/MDR also in<br />
regulation of stem cells fate. To test this hypothesis we analyzed effect of modulation of<br />
ABC-t/MDR activity in embryonic and neural stem cells. We observed that ABCC1 and<br />
ABCG2 are the most expressed ABC-t/MDR in our tested stem cells. Importantly, inhibition<br />
of these ABC-t/MDR leads to decreasing of stemness and induction of differentiation in<br />
both embryonic and neural stem cells. Analysis of mechanism of observed effect and<br />
identification of studying ABC-t/MDR substrates, which may be responsible for this<br />
effect, are in progression.<br />
Acknowledgment<br />
This work was supported by grant from MEYS CR Cost CZ LD11015 and Ministry of<br />
Education, Youth and Sport of the Czech Republic MSM0021622430<br />
P85. CYTOKINE SECRETION DURING NEURAL PROGENITOR CELL DIFFERENTIATION<br />
Karla Jarkovska 1 , Katerina Mairychova 1 , Rita Hrabakova 1 , Jirina Tyleckova 1 , Martin<br />
Marsala 2 , Silvia Marsala 2 and Hana Kovarova 1<br />
1<br />
Institute of Animal Physiology and Genetics, AS CR v.v.i., Libechov, Czech Republic<br />
2<br />
University of California, San Diego, United States of America<br />
Neural stem cells have an enormous potential in the therapy of the neurodegenerative<br />
diseases or spinal cord injuries. For therapeutic applications it is essential to characterise<br />
the developmental stages of neural cells from proliferating progenitors to fully<br />
differentiated neural cells. However, suitable biomarkers indicating commitment to a<br />
Analytical Cytometry VII 189
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