FGF2 and cyst: By starting endothelial cells induction using BMP-4 , bFGF plus Matrigel treatment, we provided a relatively closed environment for the organoid development, where the outlayer of cells are induced to form PECAM-1 positive endothelial cells, which then proliferate preferably on the Matrigel coating layer to make a cyst that prevent the inner cells for fully interact with the outside factors, allowing them to differentiate to other cell types such as neural cells.