Simplified in vitro generation of human gastruloids for modelling early development
- 1MRC Human Genetics Unit, Institute of Genetics and Cancer, University of Edinburgh, Edinburgh EH4 2XU, UK
- 2Cancer Research UK Scotland Centre, Institute of Genetics and Cancer, University of Edinburgh, Edinburgh EH4 2XU, UK
Abstract
The purpose of this study was to optimize the efficiency, cost and reproducibility of human gastruloid formation by testing and adjusting individual parameters using four human pluripotent stem cell lines available from the UK Stem Cell Bank. First, commercially sourced culture medium was replaced with a home-made defined recipe, known as ‘N2B27’, into which specific reagents can be titrated. By reducing the concentration of activin A to 15% of the original protocol, efficient elongation of aggregated embryoid bodies was achieved. Also, titrating initial cell density and delaying brief exposure to GSK3 inhibitor until the onset of cell aggregation in individual wells was advantageous. Efficiency of formation of early gastruloids exhibiting the expected regionalization of the three embryonic germ layers was further enhanced by addition of TGFβ inhibitor. The optimization steps presented thus provide a simplified, robust and relatively economical protocol for consistent generation of elongated gastruloids from human pluripotent stem cells. This streamlined method improves accessibility and reproducibility, also providing an adaptable platform to investigate fundamental principles of early human development.
Related
- Development of a simplified intensified fed-batch CHO cell culture processbased on a commercial single-feed strategy
- pSIM: A next generation in vitro mRNA potency assay.
- A quantitative pipeline for whole-mount deep imaging and multiscale analysis of gastruloids
- Development of the First Integrated Silver Nitrate-Containing In Vitro Regeneration Approach for the Endangered Tulipa montana var. chrysantha.
- Development of next-generation ALDH1B1 inhibitors with enhanced pharmacological and functional properties
BibTeX
@article{azami2026simplified,
title = {{Simplified in vitro generation of human gastruloids for modelling early development}},
author = {Takuya Azami and E. Elizabeth Patton and Jennifer Nichols},
journal = {Development (Cambridge, England)},
year = {2026},
month = oct,
doi = {10.1242/dev.205719},
url = {https://doi.org/10.1242/dev.205719},
note = {CC BY 4.0},
}
RIS
TY - JOUR
TI - Simplified in vitro generation of human gastruloids for modelling early development
AU - Takuya Azami
AU - E. Elizabeth Patton
AU - Jennifer Nichols
PY - 2026
DA - 2026/10/23
JO - Development (Cambridge, England)
DO - 10.1242/dev.205719
UR - https://doi.org/10.1242/dev.205719
AB - The purpose of this study was to optimize the efficiency, cost and reproducibility of human gastruloid formation by testing and adjusting individual parameters using four human pluripotent stem cell lines available from the UK Stem Cell Bank. First, commercially sourced culture medium was replaced with a home-made defined recipe, known as ‘N2B27’, into which specific reagents can be titrated. By reducing the concentration of activin A to 15% of the original protocol, efficient elongation of aggregated embryoid bodies was achieved. Also, titrating initial cell density and delaying brief exposure to GSK3 inhibitor until the onset of cell aggregation in individual wells was advantageous. Efficiency of formation of early gastruloids exhibiting the expected regionalization of the three embryonic germ layers was further enhanced by addition of TGFβ inhibitor. The optimization steps presented thus provide a simplified, robust and relatively economical protocol for consistent generation of elongated gastruloids from human pluripotent stem cells. This streamlined method improves accessibility and reproducibility, also providing an adaptable platform to investigate fundamental principles of early human development.
SN - 0950-1991
SN - 1477-9129
ER -