Lesson 6 Slide

Applied Molecular Cellular Biology

🧫 Organoids — Lecture Summary by Thomas Lykke-Møller Sørensen (Aarhus University)


🧬 1. Medical Biotechnology

Definition: Use of living cells or biological materials to develop therapeutic and diagnostic products that treat or prevent disease.

Examples:

  • Cell-based therapies
  • Recombinant proteins
  • Personalized medicine using patient-derived cells

🔬 2. Model Systems

Scientists use model systems to simulate human biology for study:

  • 2D Cell Culture: Simple but lacks tissue complexity
  • 3D Organoids: Mini-organs grown in vitro with realistic tissue organization
  • Animal Models: Whole-body interactions but interspecies differences exist
  • Organs-on-Chip: Mimic physiological flow and microenvironment

💡 Question on slide: “Which model systems on this line?” — invites comparing complexity and realism across these models.


🌱 3. Introduction to Organoids

Organoids are 3D multicellular structures grown from stem cells, capable of self-organization and mimicking organ function.

Key discovery: Sato et al. (Nature, 2009) – intestinal stem cells can form crypt–villus–like organoids, revolutionizing regenerative biology.


🧫 4. Organoids (Kim et al., Nat Rev Mol Cell Biol 2020)

Types:

  • Intestinal
  • Brain
  • Liver
  • Pancreatic
  • Lung

Used for:

  • Studying development, disease, drug response, and infection mechanisms.

🌱 5. Growing Organoids (Hofer & Lutolf, 2021)

Requires:

  1. Stem Cells (adult or pluripotent)
  2. Extracellular Matrix (like Matrigel)
  3. Growth Factors (Wnt, BMP, EGF, etc.)

🧩 Components:

  • Scaffold provides 3D structure
  • Signaling gradients drive regional differentiation

🧬 6. Stem Cell Differentiation

Two major paths:

  • Endoderm differentiation → gut, liver, pancreas
  • Epithelial differentiation → tissue barriers (e.g., intestinal lining)

Cells move through defined steps regulated by transcription factors and niche signals.


🧫 7. 3D Development in Matrigel

Stages (Day 0–4):

  1. Cells seeded in Matrigel
  2. Begin forming spheres
  3. Polarize and form lumen
  4. Differentiate into tissue-like structures

🧠 Matrigel acts as a “fake basement membrane” that supports 3D organization.


🔁 8. Human Organoids After 3 Passages

After several passages, organoids maintain structure and function — showing self-renewal capacity and long-term stability.


🚀 9. Intestinal Organoids 2.0

“Level-up” refers to enhancing organoid realism:

  • Adding vasculature or immune components
  • Engineering flow or mechanical stress
  • Mimicking gut microbiota

🧍‍♂️ 10. Intestinal Organoids In Vivo

Transplantation studies show:

  • Implanted organoids can integrate into damaged tissue
  • Useful for regenerative medicine

💡 11. Using Organoids – Applications

Brainstorm slide (“suggestions”): Possible uses include:

  • Disease modeling
  • Drug testing
  • Regenerative therapy
  • Infection studies
  • Personalized medicine

🧠 12. Lancaster et al., Science (2014)

Brain organoids mimicking early brain development—used to model microcephaly.


🧩 13. Gene Knockout Organoids (ACE2, TMPRSS2, Cathepsin L)

Used to study viral infection pathways (e.g., SARS-CoV-2):

  • Knockout of ACE2/TMPRSS2 → virus can’t enter
  • Cathepsin L KO → alters infection route

Shows how specific genes affect infection susceptibility.


💉 14. Therapeutic Effect of Ileum Organoid Implants

Study findings:

  • No organoid implant: rats die quickly
  • Colon organoids: partial survival
  • Ileum organoids: best survival & tissue integration

Organoid grafts can restore function after intestinal injury.


🪱 15. The Nematode Ascaris lumbricoides / suum

Parasitic worms affecting humans/pigs — model for host–parasite interactions.


✉️ 16. Extracellular Vesicles (EVs) from Ascaris suum

  • Nano-sized (30–1000 nm) membrane vesicles
  • Carry miRNA, proteins, and signals
  • Involved in cell communication and immune modulation

🔄 17. Apical-Out Phenotype

An inverted organoid structure exposing the apical surface outward, allowing:

  • Direct interaction with pathogens or compounds
  • Easier access for infection and uptake studies

🧬 18. Tuft Cells and IL-13

Tuft cells: specialized intestinal cells sensing parasites. They release IL-25, activating immune cells that produce IL-13, leading to epithelial changes and anti-parasitic defense.


🔥 19. IBD – Inflammatory Bowel Disease “To-Do List”

Goals:

  • Create patient-derived organoids from biopsies
  • Study regeneration and inflammation
  • Use bioprinting and scaffolds for reproducibility

IBD organoids enable personalized investigation of mucosal healing.


🧫 20. Colon Organoids from Patient Biopsies

Organoids derived directly from IBD patients retain:

  • Genetic background
  • Disease-specific behavior
  • Ability to test tailored treatments

🧱 21. Shaping Organoids (Mathias Lutolf’s work)

Bioengineering principles used to control shape and pattern:

  • Microwells
  • Hydrogels
  • Spatial growth factor gradients

Goal: reproducible, functional, and architecturally correct organoids.


🧩 22. Epithelial Patterning

Cells self-organize into defined epithelial zones guided by:

  • Mechanical cues
  • ECM composition
  • Gene expression domains

⚙️ 23. YAP1 Localization Window

YAP1 (mechanosensor protein) determines if cells proliferate or differentiate:

  • Cytoplasmic → quiescent
  • Nuclear → active growth There’s a “window of opportunity” where YAP1 localization determines fate.

🧠 24. Bioengineered Organoids

Combining stem cells with:

  • Synthetic matrices
  • Microfluidics
  • Controlled signaling environments

Leads to organoids with improved reproducibility and function.


🧩 25. New Organic Designs

Innovative engineering approaches producing customized mini-organs with defined shape and function (e.g., gut-on-chip, brain chips).


📊 26. Quantification in ImageJ

ImageJ used for:

  • Measuring organoid size, shape, and growth
  • Quantifying cell density and fluorescence Essential for objective comparisons.

⚖️ 27. Ethical Issues (Q3)

Prompt for reflection:

  • Brain organoids with neural activity → consciousness concerns?
  • Genetic modification ethics
  • Use of human cells and patient consent

🧠 28. Brain Organoids

Mimic human brain development and function in 3D. Used to study:

  • Neurodevelopmental disorders
  • Virus infections (e.g., Zika)
  • Tumor biology

🧩 29. Cancer Organoids

Patient-derived tumor organoids allow:

  • Personalized drug screening
  • Genotype–phenotype studies
  • Testing drug resistance (Jacob et al., Cell 2020)

🧠 30. Brain Organoids (Lancaster et al., Nature 2013)

Show early brain patterning, neural tube-like regions, and cortical organization.


🔄 31. Differentiation in Brain Organoids

hiPSCs → embryoid bodies → neural induction → cortical tissue layers. Recapitulates early neurogenesis.


⚗️ 32. Growth of Cerebral Organoids

Challenges:

  • Central necrosis due to lack of vasculature
  • Variable embryoid body size
  • Solutions: smaller seeding pools, separated culture wells

📈 33. Growth Curve and Size Differences

Quantitative imaging shows growth variability; standardization needed for reproducible results.


🧩 34. Connected Organoids on PDMS-MEA Chip

Two brain organoids connected via microelectrode array (MEA) → record neuronal communication and network activity — “mini-brains” interacting!


⚡ 35. Generating Complex Neuronal Activity

Connected organoids show spontaneous electrical signaling, similar to neural circuits — foundation for studying learning-like processes in vitro.


🧠 36. Axon Bundle Population

Formation of axon tracts linking two organoids — showing real synaptic connectivity between separate “brain islands.”


🧩 37. Connecting Organoids (Anne Louise Jackson, MSc Thesis)

Used GelMA hydrogels and unidirectional scaffolds for guided neural outgrowth between organoids. Stained for:

  • F-actin → cytoskeleton
  • βIII-tubulin → neurons

Demonstrates engineered connectivity in vitro.


🧠✨ Summary Takeaways

ConceptKey Idea
Organoids3D self-organizing stem cell–derived mini-organs
ApplicationsDisease modeling, drug screening, regenerative therapy
Engineering advancesControlled shape, bioengineered scaffolds, vascularization
Ethical issuesEspecially relevant for brain organoids
Future outlookTowards functional, connected, and patient-specific organoid systems

Quiz

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