Combination of epigenetic erasing and mechanical cues to generate human epiBlastoids from adult dermal fibroblasts
Articolo
Data di Pubblicazione:
2023
Citazione:
Combination of epigenetic erasing and mechanical cues to generate human epiBlastoids from adult dermal fibroblasts / G. Pennarossa, S. Arcuri, T. De Iorio, S. Ledda, F. Gandolfi, T. Brevini. - In: JOURNAL OF ASSISTED REPRODUCTION AND GENETICS. - ISSN 1573-7330. - 40:5(2023 May), pp. 1015-1027. [10.1007/s10815-023-02773-4]
Abstract:
PurposeThis study is to develop a new protocol that combines the use of epigenetic cues and mechanical stimuli to assemble 3D spherical structures, arbitrarily defined "epiBlastoids," whose phenotype is remarkably similar to natural embryos.MethodsA 3-step approach is used to generate epiBlastoids. In the first step, adult dermal fibroblasts are converted into trophoblast (TR)-like cells, combining the use of 5-azacytidine, to erase the original phenotype, with an ad hoc induction protocol, to drive cells towards TR lineage. In the second step, epigenetic erasing is applied once again, in combination with mechanosensing-related cues, to generate inner cell mass (ICM)-like organoids. Specifically, erased cells are encapsulated into micro-bioreactors to promote 3D cell rearrangement and boost pluripotency. In the third step, TR-like cells are co-cultured with ICM-like spheroids in the same micro-bioreactors. Subsequently, the newly generated embryoids are transferred to microwells to favor epiBlastoid formation.ResultsAdult dermal fibroblasts are successfully readdressed towards TR lineage. Cells subjected to epigenetic erasing and encapsulated into micro-bioreactors rearrange in 3D ICM-like structures. Co-culture of TR-like cells and ICM-like spheroids into micro-bioreactors and microwells induces the formation of single structures with uniform shape reminiscent in vivo embryos. CDX2(+) cells localized in the out layer of the spheroids, while OCT4(+) cells in the inner of the structures. TROP2(+) cells display YAP nuclear accumulation and actively transcribed for mature TR markers, while TROP2(-) cells showed YAP cytoplasmic compartmentalization and expressed pluripotency-related genes.ConclusionWe describe the generation of epiBlastoids that may find useful application in the assisted reproduction field.
Tipologia IRIS:
01 - Articolo su periodico
Keywords:
3D culture systems; Epigenetic erasing; ICM-like cells; Mechanosensing-related cues; TR-like cells; epiBlastoids;
Elenco autori:
G. Pennarossa, S. Arcuri, T. De Iorio, S. Ledda, F. Gandolfi, T. Brevini
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