Differentiation of mouse P19 embryonic carcinoma stem cells injected into an empty zebrafish egg chorion in a microfluidic device

Mouse P19 embryonic carcinoma (EC) stem cells were xenotransplanted into the emptied chorion, the transparent envelope of a fertilized zebrafish egg (rather than mouse native zona pellucida) combined with a microfluidic device to study P19 EC cell differentiation in the chorion biomaterial. A distilled-water jet was used to remove the innate yolk and perivitelline inner mass from the chorion. P19 EC cells were injected into the emptied chorion using a micropipette, and they were subsequently cultured until the inner space of the chorion became completely occupied by cells. A simple microfluidic device was used for handling convenience and effective experiment. At d15, we found neural cells in the outer layer of the cell mass and beating cardiomyocytes in the inner layer of the large embryoid body. We propose that even though the species are different, the external innate membranes developed for embryo protection represent a useful type of ECM.