Lancaster et al., Nature 2013

GROUP 1 (Matteo Preguglio Mulè Francesca, Celeste Nicola, Dondu Bozkurt)

- What are embryoid bodies?

- What is the origin of the cells used in the experiment represented in fig. 1?

- On what bases the organoids were identified as cerebral? 

- On what bases different brain structures were identified to be part of the brain organoids? Is the hippocampus formed in brain organoids? 

- How big (n?) is the sample analysed? Can you find this information in the paper?

-In the study the authors show that similar cerebral organoids can be derived starting from hES and iPS cells – what do you think is the relevance of this observation?

Fig.1 – Explain what is the main message given and what kind techniques were used to visualize specimens 

Fig. 2 – Explain the figure (what is the main message) - do you think the information given in this figure is exaustive?

In the legend: What does it mean serial section? What is tangential migration?

Based on their observations, authors suggested that cell migration occurs in organoids (i.e. cortical interneuron migration from ventral to dorsal forebrain – what experiment do you think can be designed to test their suggestion?

GROUP 2 (Marta Di Renzo, Giulia Zuccarini, Daliene Stajano, Fabiana Rossi)

- Compare the two sections: Recapitulation of dorsal cortical organization (fig.3) and Recapitulation of human cortical organization (fig.5). Describe the figures and the main message.

What are RGs and oRGs?

What is interkinetic nuclear migration?

Why p-vimentin immunostaining is used to examine division plane in RGs?

How do they test if RGs were dividing simmetrically or asymmetrically?

Why they used in vitro electroporation following injection of GFP plasmids in brain organoids? 

The study is centered on human brain development, why for some experiments mouse-derived brain organoids were introduced?

GROUP 3 (Marco Ghibaudi, Viviana Canicatti, Filippo Michelon)

Cerebral organoids model human microcephaly -Refer to Figure 6 and extended fig7 and 8

What is the rationale of the experiment?

Which experimental approaches were used to test the implication of CDK5RAP2 in brain development?

What were the parameters analysed to check for abnormal development?

GENERAL

What are the main limits encountered in growing cerebral organoids?

Based on what you read in this paper is there a large or small sample-to sample variability?

What is a spinning bioreactor? What are the major advantages compared to static culture and what could be the major limits in using this technology?

Do you think the brain organoids described in this study could represent a useful tool to investigate how the 6 layered structure of the cerebral cortex is formed? 

In which direction do you think researchers must work for further development of this technique?