MF9430 – MicroRNA and transcription factors as determinants of cell phenotypes and stem cell differentiation
The course describes microRNA and transcription factor (TF) interactions as a key regulatory system in the determination of cell phenotype through regulation of marker gene expression. It also describes how microRNA signatures of cell phenotypes are determinant for normal organ functioning and pathophysiology.
Practical laboratory course and bioinformatics analyses. The objective is to provide practical information on how microRNA and transcription factors (TFs) interact in differentiating stem cells, how to use a microRNA species as disease markers. Furthermore, different databases (including Mir@nt@n)) will be used as part of the strategy to identify important microRNA-vs-TF networks. Lastly, the students will get some hands on experience related to the anlyses of microRNA and TF expression, as well as traditional staining techniques identifying osteoblasts, chondrocytes and adipocytes differentiated from mesenchymal stem cells or fat tissue derived stem cells.
Ph.D. candidates and students at the Medical Student Research Programme will get first priority to the course. The maximum number of participant is 30.
Formal prerequisite knowledge
Cellbiology, Biochemistry, Physiology, Embryology etc.
Recommended previous knowledge
Generegulation, signaling pathways
• Introduction to course: Differentiation of stem cells to osteoblasts, chondrocytes and adipocytes
• Transfection of siRNAs to suppress mRNAs of transcriptionfactors important for osteoblast, chondrocyte and adipocyte differentiation on plated stem cells. Incubation for 3 days.
• Cell staining techniques, light microscopy, assessment of percentage of marker-positive surfaces (mineralization, alkaline phosphatase, proteoglycans, fat droplets) using predifferentiated osteoblasts, chondrocytes and adipocytes from 14 day cultures
• Isolation of total RNA from 3 days differentiated osteoblasts, chondrocytes, and adipocytes. Q-PCR analyses of transcription factors (TFs) and selected microRNAs
Grading: Will be based on seminar performances, demonstration of practical skills, an essay with title/subject chosen by the participant, and the results obtained in the “multiple choice” session.
The examination papers will be scanned through the ephorus program.
Explanations and appeals
The course is subject to continuous evaluation. At regular intervals we also ask students to participate in a more comprehensive evaluation.