A Technique for Generating Neural Progenitor Cells from Mouse Embryonic Stem Cells
Transcrição
Take mouse embryonic stem cells, or mESCs, suspended in a differentiation medium, and incubate.
In the suspension culture, mESCs aggregate to form a structure termed embryoid bodies or EBs.
EBs are three-dimensional aggregates that mimic a developing embryo and can be differentiated into neural cells.
Transfer the EBs into a tube and allow them to settle by gravity.
Remove the nutrient-depleted medium; resuspend the EBs in a fresh medium, and transfer them to a culture dish.
Incubate the suspension culture to facilitate the growth of EBs.
Transfer the EBs suspended in the medium onto a multiwell plate. The wells are coated with an adhesion protein, allowing the EBs to attach.
Introduce retinoic acid, which enters the cells and triggers the formation of a transcription complex.
The complex induces gene expression, promoting the differentiation of the cells into neural progenitor cells, or NPCs, which exhibit extended cellular processes.
