Organoids
Agnieszka Rybak-Wolf
Profil
The organs of the human body consist of distinct cell types that are organized in a specific manner to form a multi-layer network. Today it is possible to reconstruct organ-like tissues (organoids) in the laboratory, whereby stem cells are directed by molecular signals that drive their differentiation and are grown in culture systems that promote their three-dimensional self-organization. Organoid technologies, which are rapidly developing, allow to phenotypically copy the cellular composition and, to a certain degree, also the functionality of various human organs such as brain, thyroid, thymus, intestine, liver, pancreas, stomach, lungs, kidneys, and even early embryos. As near-physiological 3D culture systems, organoids open up new possibilities for investigating the development of healthy and diseased organs and offer great potential for translational research.
"Throughout my scientific career, I have been interested in how post-transcriptional gene regulation drives neuronal differentiation and brain development. The focus of my research concerns regulatory RNAs, such as miRNAs and circular RNAs, and their expression and function in the healthy and diseased mammalian nervous system. As leader of the newly established organoid platform at MDC, I aim to develop 3D brain models for diseases such as Leigh Syndrome, Alzheimer’s disease and infectious brain diseases, and work to elucidate the molecular mechanisms underlying disease development and progression. A major goal of the platform is to further engineer the brain organoid culture systems and improve in-vitro neural tissue maturation. We currently implement technologies to vascularize brain organoids (assembly of brain and blood vessel organoids) and incorporate microglia into our systems in order to realize long-term maturation of three-dimensional neural tissues. I’m a mother of two creative and very energetic twins." - Dr. Agnieszka Rybak-Wolf
Gallery of silly organoids (use your imagination)
Team
“The centre of my current research interest is optimization and development of brain tissue models for basic and clinical research and in particular for studying mechanisms of neurodegenerative disorders” Agnieszka Rybak-Wolf
“Here at the organoid technology platform, my motivation revolves around 2 key aspects: In general, I want to improve the translatability of organoid systems for biomedical research and real-life applications. From a more applied perspective, I aim at exploiting organoid technology to study age-related diseases” Nicolai Kastelic
“After studying Neuroscience at the University of California, Berkeley and at Albert-Ludwigs-Universität Freiburg, I've had the opportunity to participate in many diverse projects. In my decade of lab work, I've studied neuro-degeneration in cell culture, been a honeybee brain surgeon, and used deep learning to listen in on the secret vocalizations of mice, to name a few highlights. I am currently interested in neurodiveristy and the mechanisms of executive dysfunction. I am delighted to join the Organoid Platform and look forward to innovating exciting new models’’ Josephine Coburn
Former lab members
Dr Anna Loewa
Ruth Pareja
Technology & Application
Engineering brain organoid culture systems
The Organoid Platform was established in 2019 as a part of the Berlin Institute for Medical Systems Biology (BIMSB) at the Max Delbrück Center (MDC). The mission of the platform is to support organoid-related projects at the MDC by establishing state-of-the-art organoid protocols, implementing those protocols within collaborative projects with MDC research groups, providing training and developing new methodologies. In order to address the needs of MDC researchers, we currently focus primarily on brain tissue models, but the range of mini-organs is expanding and will be further expanded in the coming years.
Our current goal is to further engineer brain organoid culture systems and to improve in vitro neural tissue morphogenesis. We are now implementing technologies to vascularize brain organoids and to incorporate microglia (a type of non-neuronal cells in the central nervous system) into our systems in order to realize long-term maturation of three-dimensional neural tissues. In addition, we set up methods for organoid characterization including single-cell and spatial transcriptomic technologies, electrophysiological measurements, immunostainings and live imaging.
A collaborative effort to dissect mechanisms of disease
In addition, we collaborate with Pluripotent Stem cell platform and several MDC groups, implementing brain organoids to model human brain diseases such as:
- Leigh Syndrome - PMCID: PMC7997884,
- Herpes simplex encephalitis (HSE) - doi: https://doi.org/10.1101/2021.03.05.434122,
- Brain tumors (manuscript in preparation)
and to test new therapeutic approaches to improve the current treatment of these diseases.
Our ultimate goal is to create more complex brain tissue models addressing experimental and translational needs and to create a general blueprint for modern therapies in which personalized patient-derived brain organoids serve for in vitro disease modeling and drug screening to improve current therapies.
Collaborative Research Project
Highlights
| Project |
Result/acknowledgement/publication |
|
| Finished projects | ||
| 1. | FLAMseq |
Legnini et al., 2019 Platform acknowledgment |
| 2. |
Leigh Syndrome |
Inak et al., 2021, Nat Comm and Le S et al., 2021 |
| 3. |
Modeling HSV-1 driven encephalitis in brain organoids |
Rybak-Wolf et al., (accepted, Nature Microbiology) |
| 4. |
Organoid patterning via optogenetic activation |
Legnini et al., accepted, Nature methods |
| 5. |
A circular RNA expressed from the FAT3 locus regulates neural development |
Seeler et al.,2023 Mol Neurobiol |
| 6. |
Sites of transcription initiation drive mRNA isoform selection |
V.Hilgers, N.Rajewsky |
| 7. |
circRNA expression and function in glioblastoma |
manuscript in preparation |
| 8. |
Huntington project |
manuscript in preparation |
Third Party Funding
2020 BIH Platform Humanized Model Systems
‘’Assembloid of brain organoid and patient-derived neurosphere as a physiological model for glioblastoma invasion study’’ in collaboration with K.Rolle (ICHBPAN)
2023 MDC Tandem grant
‘’Streamlined active labeling for 3D protein visualization in intact cleared organoids’’ in collaboration with R. Zinzen (MDC)
2023 MDC Tandem grant
“A matrigel-free method to generate mature human brain organoids using celvivo bioreactor system’’ in collaboration with N.Rajewsky (MDC)
2023 Charite 3R call
co-applicant, ‘’Modeling defective Ras signaling in human cerebral organoids – Towards a therapy for the severe neurodevelopmental RASopathy SynGAP syndrome” project lead Sarah Shoichet (MDC)
Publications
News
Models
- Cerebral, forebrain and midbrain organoids
(Lancaster et al., 2014; PMID: 25188634, Quian et al., 2016; PMID: 27118425) - ALI/enCOR organoids
(Giandomenico et al, 2019; PMID: 30886407, Lancaster 2017; PMID: 28562594) - Assembloids
(Bagley et al., 2017, PMID: 28504681) - Bioengineered Neuronal Organoids (BENOs)
(Zafeiriou et al., 2020; PMID: 32728089) - 3D silk collagen brain tissue models
(Chwalek et al., 2015, PMID: 26270395) - Blood-vessel organoids
DOI : https://doi.org/10.1038/s41586-018-0858-8 - Tumoroids
PMID: 31883794
Services
- Training iPSC/NPCs culture/organoid culture
- Organoids derivation
- platform iPSC lines
- custom lines - ClinoStar bioreactor organoid culture
- Organoids vascularization and microglia integration
- Organoid transgenesis (NEPA/AMAXA electroporation, lipofection, viral transduction)
- Organoid characterization:
- nanostring (standard or custom panel)
- imunohistochemistry/ISH
- MEA (Axion BioSystems, 3Brain) - Organoid preparation for scRNAseq and spatial transcriptomic
- Disease modeling
- Gene editing/clonal expansion
