Figure 2
From: Engineered 3D vascular and neuronal networks in a microfluidic platform
MN spheroid formation and characterization and morphogenesis of NSC-MN and MNP-MN spheroids. (a) The diameter of MN spheroids can be controlled by changing the seeding density. Immunostaining of Tuj1(green) and DAPI (blue) in NSC-MN and MNP spheroid. Left: NSC-MN spheroid. Right: MNP-MN spheroid. (b) Comparison of relative gene expression change of neuronal markers (Oligo2, HB9, ISL1, and GFAP) between NSC-MN spheroids and MNP-MN spheroids. NSC-MN spheroids have higher expression the gene related mature MN (islet1). In addition, the expression of GFAP which is astrocyte marker can be detected in NSC-MN spheroids. n = 4; *P < 0.01, two-way ANOVA. (c,d) Relative gene expression change of MN markers (Oligo2, HB9, islet1) and Nanog and neuronal stem cell marker (Nestin) showed that spheroid culture significantly improves differentiation into mature MNs compared to traditional monolayer culture. n = 5; *P < 0.01, two-way ANOVA. (e) No significant difference of hypoxia marker (HIF-1α) between the spheroid and monolayer culture. n = 3; *P < 0.01, two-way ANOVA, error bars ± SD.
