Abstract: The present disclosure provides a bio-ink cartridge, a bio-ink cartridge assembly, a microsphere preparation device, a shell assembly device, a bio-block preparation instrument, a bio-ink preparation instrument and a bio-ink preparation system. The bio-ink cartridge comprises an ink cartridge shell with a cavity; a dividing wall arranged in the cavity and dividing the cavity into a first accommodating chamber and a second accommodating chamber independently; a first ink cartridge inlet and a first ink cartridge outlet which are arranged on the ink cartridge shell, communicated with the first accommodating chamber and separated from the second accommodating chamber; a second ink cartridge inlet and a second ink cartridge outlet which are arranged on the ink cartridge shell and communicated with the second accommodating chamber; and an inner flow channel located in the second accommodating chamber and communicated with the first accommodating chamber and the first ink cartridge outlet.