sustained synchronized network activity was achieved, starting three weeks after final plating of neurons (~DIV50) and sustainable up to 8 weeks (~DIV90) in vitro, with neglectable variation between two different hiPSC lines. The support of primary human astrocytes, via released growth factors and physical contact31, in combination with forced (DAPT) simultaneous differentiation of NPCs into neurons probably leads to more homogenous and mature networks in a shorter time frame. This way, there is no need to wait for the delayed emergence and development of astrocytes from NPCs, which causes slow but progressive maturation of the network44.
