Figure 6
From: Engineered 3D vascular and neuronal networks in a microfluidic platform
Perfusion culture influences synaptogenesis of MN networks along with synapse elimination. (a) Formation of hiPS-EC networks (cyan) and MN networks (purple) in a microfluidic device in the presence and absence of perfusion culture after 7 days of culture. There is no significant difference of formation of microvascular networks. Dextran perfusion demonstrated patent endothelial networks in both conditions. (b,c) Ca2+ imaging demonstrated that perfusion culture suppresses neurite extension compared to static culture quantified by neurite tracking (purple). (d) Spontaneous Ca2+ oscillation showed perfusion culture upregulated neural activity and synaptic connections, whereas total neurite length is shorter than static culture on day 7. (e) The representative waveform of Ca2+ imaging indicated that MN networks with perfusion culture higher response of depolarization of action potential than static culture. (f,g) Quantification of spike frequency and peak calcium concentration showed perfusion culture significantly accelerated synaptogenesis along with synapse elimination. n = 3; *P < 0.01, **P < 0.05, student’s t-test. Error bar ± SD.
