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Digital archive of theses discussed at the University of Pisa

 

Thesis etd-05192021-124208


Thesis type
Tesi di laurea magistrale LM6
Author
BARBIERO, SILVIA
URN
etd-05192021-124208
Thesis title
Distinct metabolic states sustain the emergence of cell lineages in intestinal organoid regeneration
Department
RICERCA TRASLAZIONALE E DELLE NUOVE TECNOLOGIE IN MEDICINA E CHIRURGIA
Course of study
MEDICINA E CHIRURGIA
Supervisors
relatore Prof.ssa Brunetto, Maurizia Rossana
correlatore Prof.ssa Liberali, Prisca
Keywords
  • intestinal organoid
  • metabolism
  • regeneration
  • single-cell decision-making
Graduation session start date
15/06/2021
Availability
Withheld
Release date
15/06/2091
Summary
From the smallest cell to the whole body, thousands of gears work in concert in space and time to ensure the utmost expression of human physiology. How in the absence of a hierarchical control, single cells coordinate and give rise to complex tissues is a mystery of multicellular organisms. Borrowing from the science of complex systems, it has been hypothesized that a set of local rules govern the life of a single cell and that emergent properties stem from complex interactions at the population level. Here we study single-cell decision-making in the context of a complex self-organising eukaryotic tissue.
With one of the highest turnover rates in the body, intestinal epithelium is a flourishing and dynamical territory where regenerative cell-types constantly sense the environment and need to decide which fate to acquire and which mature cell-type to become. To quantitatively track the single cells whilst keeping a holistic view of the emergent patterns at the population levels, we made use of organoid techniques which allow the in-vitro three-dimensional reproduction of the microanatomy and functions observed in-vivo. We studied the process of intestinal organoid regeneration, whereby a single cell rapidly grows into a symmetrical sphere of cells which ultimately buds into an asymmetrical mature organoid, with its crypts and villi that morphologically and functionally recapitulate the intestinal epithelium.
Through the transcriptomic analysis at the single cell resolution, we characterized the metabolic fingerprints that support intestinal organoid regeneration and cell lineage acquisition. We further confirmed our results through high-throughput imaging in fixed and living single cells from mature intestinal organoids. Through manipulation experiments targeting newly found metabolic markers we generated organoids with different proportions of mature cell-types. Altogether, our findings suggest an instructive role for metabolism in cell decision-making, suggesting cell metabolic state as a putative candidate for understanding the coordination and collective behaviour at the system level.
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