Course Syllabus
Microbrains Lab: Modeling Neurodegeneration |
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| Semester & Location: |
Summer 2022 - DIS Copenhagen |
| Type & Credits: |
Summer Lab course - 6 credits |
| Major Disciplines: |
Biology, Biomedicine / Biotechnology, Pre-Medicine / Health Science |
| Prerequisites: |
One year each of biology and one course in either molecular biology or genetics, all at university level. |
| Faculty Member: |
Kristine Freude |
| Program Director: | Susana Dietrich |
| Time & Place: |
DIS: Lab work: Look at syllabus for details |
Faculty
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Kristine Freude, Associate Professor PhD at Max Planck Institute for Molecular Genetics and Free University, Berlin, Germany in Human Genetics (2005). Postdoctoral Research Fellow at UC Irvine, CA, USA (2005-2011) Postdoctoral Researcher (2012-2014) University Copenhagen. MSc at Robert Koch Institute and Free University, Berlin, Germany, in 2001 working with Mycobacteria. Assistant Professor (2014-2015) University Copenhagen. Associate Professor (2015-present) University Copenhagen. With DIS since 2016. |
Course Description
Stem Cells and especially human induced pluripotent stem cells (hiPSC) have been instrumental for generating human in vitro models for both better understanding and drug screening for neurodegenerative diseases. In this course you will learn to culturing of hiPSC and genetic modifications of hiPSC implementing CRISPR-Cas9 precise genome editing. Moreover you will learn how to learn how to differentiate those hiPSC into neurons, astrocytes and microglia as well as 3D retinal organoids. In addition to the hands on lab work you will get the theoretic background and you will be introduced to the latest research within the field.
Expected Learning Outcomes
- To understand the concepts of pluripotent cells and their potential.
- To learn to culture and differentiate iPSC into various brain cells and 3D organoids
- To be able to write a concise lab report
- To be able to present, interpret and analyse scientific articles related to the course and present them to your peers.
Description of Assignments, Testing and Grading
Evaluation
To be eligible for a passing grade in this class all of the assigned work must be completed.
The factors influencing the final grade and their weights are reported in the following table:
| Class participation | 20% |
| Oral presentations at journal clubs | 20% |
| Lab Report | 50% |
| Attendance | 10% |
| Total | 100% |
Class participation covers the following areas:
- Level of preparation
- Contribution to class discussions & journal clubs
Class participation is an important part of this course and to receive full credit students should be present at all the scheduled classes and actively participate.
Journal Clubs
Journal clubs are presentations followed by discussions that will be organized throughout the course to analyze certain thematic areas in depth.
Journal clubs are occasions for group work, class discussion, and development of presentation skills; furthermore, they serve as an exercise for critical thinking and reading.
The papers presented during the Journal Club are related to the preceding lesson. The presenting group of the week will have to prepare a presentation. The rest of the class should read the paper as well and prepare one or two questions for the class discussion.
The presentation should have a logical and clear structure and provide relevant information on the background, methods, conclusions, and future perspectives of the presented work. The original data reported in the paper should be presented and discussed in a clear way.
Lab Report
As part of the course you will prepare a Lab Report. The purpose of the Lab Report is to learn and practice how to document you research experiments and protocols, that everyone can pick up your report and recapitulate your experiments and results. Moreover you should critically evaluate your results and elaborate on possible improvements of your methodology and discuss your results. Further descriptions and instructions will be given during the course.
Required Readings
- Material will be posted on Canvas in the relevant modules. It is expected that you read the relevant material ahead of the module/week.
Policies
Laptop policy
Use of laptop computers is not allowed in the laboratories, but encouraged in the class rooms. Cell phones and other electronic devices should be turned off and stored away.
DIS Contacts
Susana Dietrich, Program Director, shsupport@dis.dk
Science & Health Program Office: Vestergade 7-37
Academic Regulations
Please make sure to read the Academic Regulations on the DIS website. There you will find regulations on:
Overview of course's workload distribution
Week 1: Basics in stem cell research
Theoretical part:
- Lectures on embryonic stem cells, induced pluripotent stem cells, reprogramming methods and ethical considerations.
- Journal club performed by students (topic hiPSC and ESC)
- Introduction into lab reporting and daily supervision and reflections to perform the final report
Practical Part:
- Thawing and expanding human induced pluripotent stem cells (hiPSC)
- RNA extraction
- qPCR to assess pluripotency
- immunocytochemistry for pluripotency proteins
- Freezing of hiPSC
Week 2: Embryonic development and lineage commitment
Theoretical part instructor:
- Lectures on embryonic development, early lineage commitment, brain development
Theoretical exercises students:
- Journal club performed by students (topic brain development)
- Continuous work on lab reports
Wet lab exercises students:
Induction into neuronal linages
- Continuation of experiments from week one with microscopic analyses of ICC and calculation and statistics for qPCR.
Week 3: 2D neurodegenerative disease models, including CRISPR
Theoretical part instructor:
- Lectures on neurodegenerative diseases and how to model those in vitro, CRISPR-Cas9 basics and implications in disease modeling
Theoretical exercises students:
- Journal club performed by students (neurodegenerative diseases and CRISPR-Cas9)
- Continuous work on lab reports
Wet lab exercises students:
- Analyses of neural progenitors from week 2, ICC, qPCR and morphology
- CRISPR-Cas9 gene editing of hiPSC and clonal selection over the last three weeks.
Week 4: Differentiation into neuron subtypes
Theoretical part instructor:
- Lectures on neurodegenerative diseases and subtypes of neurons affected.
Theoretical exercises students:
- Journal club performed by students (different approaches to get specific neural subtypes)
- Preparation by students of protocols for neural differentiation
- Continuous work on lab reports
Wet lab exercises students:
- Maturation of NPCs into glutamatergic and GABAergic neurons
- Clonal selection of CRISPR reprogrammed hiPSC
Week 5: Differentiation into glia
Theoretical part instructor:
Lectures on the role of astrocytes and microglia in neurodegenerative diseases.
Theoretical exercises students:
- Journal club performed by students (different approaches to get microglia and astrocytes and relevance in disease context)
- Preparation by students of protocols for astrocyte and microglia differentiation
- Continuous work on lab reports
Wet lab exercises students:
- Continuous maturation of NPCs into glutamatergic and GABAergic neurons
- Continuous clonal expansion of CRISPR reprogrammed hiPSC
- Induction of macrophage progenitors and astrocyte progenitors into microglia and astrocytes respectively.
Week 6: Organoid models for neurodegenerative disease
Theoretical part instructor:
- Lectures organoid models to study neurodegenerative diseases.
Theoretical exercises students:
- Journal club performed by students (organoids – cerebral, thalamic, retinal etc.)
- Preparation by students of protocols for organoid differentiation
- Continuous work on lab reports
Wet lab exercises students:
- Assessment of glutamatergic and GABAergic neurons via ICC and qPCR
- Analyses of selected CRISPR clones for successful integration
- Establishment of 3D cultured retinal organoids from 2D cultures.
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