Description
Cooperation is focused on biomedical education and stem cell biology; it is aimed at deepening the bilateral relationship by promoting and intensifying contacts and mobility between both participating partners. Interactions involve practical training in the labs, extending the knowledge and practice in cell reprogramming, cell manipulation, characterization, directed differentiation, formation of 3D organoids and also mutual discussions. We will carry out optimization of array cards for gene expression in iPSCs and analysis of isolated cell clones incl. histological examination of generated organoids. Partner teams include academic workers, pregradual and postgradual students. Seminars and lectures will be organized to involve broader spectrum of investigators from related disciplines to establish further future cooperation and get students the opportunity to obtain detailed information directly from prominent experts and have discussions with them.
Summary of project results
Induced pluripotent stem cells (iPSCs) are genetically reprogrammed (in our set-up human) somatic cells exhibiting a pluripotent stem cell-like state similar to embryonic stem cells. Thus, they are an invaluable new source of pluripotent cells for drug discovery, cell therapy and basic research. Cellular reprogramming technology and personalized medicine with regard to modelling diseases poses promising ad hoc utilization of iPSCs in a clinical approach with focus on studies that have demonstrated a disease phenotype in the tissue of interest. The iPSCs PCR array card should profile the expression of the key genes involved in iPSC research. To control the procedure of cell reprogramming and conversion to pluripotency, the expression of multiple gene classes included on the array has to be monitored simultaneously: representative parental cell line genes, the ectopically expressed transcription factors, markers of iPSCs, and markers of the redifferentiation into cells of ectodermal, endodermal, and mesodermal origin. Namely, we can “functionally” divide tested genes into 5 groups: expression in undifferentiated cell, maintenance of pluripotency, correlation to stemness, differentiation markers, endogenous controls - housekeeping genes.Using real-time PCR, research studies can easily and reliably analyse the expression of a focused panel of genes involved in the induced pluripotent stem cell dedifferentiation and redifferentiation processes with this array.
Summary of bilateral results
The knowledge of our NP was utilised for the generation of iPSCs and their propagation. In addition, our partner provided guidance concerning characterisation and directed differentiation toward desired cell types, techniques routinely managed in their lab. Ongoing establishing of cellular reprogramming technology in our laboratory was focused on the generation of reprogrammed cells derived from patients. Long-term goal of collaboration will be to participate on the preparation of disease model for drug discovery and compound screening. Using real-time PCR we could reliably analyse the expression profiles of a focused panel of genes involved in the induced pluripotent stem cell dedifferentiation and redifferentiation processes with the array. We evaluated differences of gene expression in parental cells in comparison to reprogrammed ones to distinguish both successfully reprogrammed cells and to understand the process on the molecular level. Within this framework following activities were realised; mobility of two academics and four students from the Czech Republic to Norway, in order of laboratory training and optimising IPSCs array cards. Three academics and one student from Norway visited the Czech Republic. Dr. Sullivan held a lecture in Prague and all of them including Czech partners participated on the two-day workshop in Hradec Kralove. Norwegian partner received 200 iPSCs array cards, which were modified based of previous tests when the set of genes for testing individual patient-specific iPSCs clones was optimised, and probes for six “naive” genes were added. Establishing cellular reprogramming technology in our laboratory and the complex testing of iPSCs array card are main results of our mutual collaboration.