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| Status |
Public on Jun 12, 2012 |
| Title |
Confined 3D microenvironment regulates early differentiation in Human Pluripotent Stem Cells |
| Organism |
Homo sapiens |
| Experiment type |
Expression profiling by array
|
| Summary |
We aimed to investigate how different three-dimensional microenvironments regulate the early differentiation of the three germ layers in human embryonic stem cells derived embryoid bodies. In particular, a permeable, biocompatible, hydrogel microwell array was specifically designed for recreating a confined niche in which EB secreted molecules accumulate in accordance with hydrogel diffusional cut-off. Fluorescence recovery after photobleaching technique was performed to accurately evaluate hydrogel permeability, mesh size and diffusional cutoff for soluble molecules. EBs culture in microwells promotes the expression of genes involved in pattern specification processes, brain development, ectoderm and endoderm differentiation. On the contrary, suspension EBs express instead genes involved in mesoderm specification and heart development. These results suggest that local accumulation of EBs secreted molecules drives differentiation patterns, as confirmed by immunofluorescence of germ layer markers, in hydrogel confined EB culture.
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|
| Overall design |
Three different culture conditions of EB culture were analyzed: suspension (standard condition), confinement in microwells of width/depth ratio 1:1 and 1:2. EBs cultured in microwells are viable and have comparable average size after 8 days culture. Whole genome microarrays show that significative differential gene expression was observed between suspension and confined EBs culture.
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| |
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| Contributor(s) |
Giobbe GG, Zagallo M, Riello M, Serena E, Masi G, Barzon L, Di Camillo B, Elvassore N |
| Citation(s) |
22674472 |
| Submission date |
Mar 13, 2012 |
| Last update date |
Jan 23, 2019 |
| Contact name |
Barbara Di Camillo |
| E-mail(s) |
barbara.dicamillo@dei.unipd.it
|
| Organization name |
University of Padova
|
| Department |
Information Engineering Department
|
| Street address |
via Gradenigo 6B
|
| City |
Padova |
| ZIP/Postal code |
35125 |
| Country |
Italy |
| |
|
| Platforms (1) |
| GPL6480 |
Agilent-014850 Whole Human Genome Microarray 4x44K G4112F (Probe Name version) |
|
| Samples (9)
|
| GSM894482 |
Hydrogel_1:2ratio(width/depth)_8days_rep1 |
| GSM894483 |
Hydrogel_1:2ratio(width/depth)_8days_rep2 |
| GSM894484 |
Hydrogel_1:2ratio(width/depth)_8days_rep3 |
|
| Relations |
| BioProject |
PRJNA153515 |
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