Fig. 5.

Characterization of early-stage maturation of organoids. A) Immunocytochemistry of actin in LRC, HRC, HRF and CON revealed detailed lumen structures and cell organization, as outlined by white dashed lines. B) The diameter of lumens on day 20 was significantly larger in LRC and HRC group than CON, with the HRC group having the largest lumens. C) The thickness of lumens on day 20 shared a similar trend with the lumen diameter. D) The number of lumens on day 20 was significantly larger in all microwell groups than CON, while the HRC group generated the highest lumen numbers. E) Immunocytochemistry of organoid cryogenic sections on day 45 revealed neuronal cell layers, as labeled by mature neuron related markers NeuN and Map2. F) The thickness of Map2 positive cells in CON, HRF, LRC and HRC averaged over at least 3 organoids. The thickness of the mature layer was significantly increased in LRC and HRC group. The HRC group has the maximum average value of the thickness of mature neurons. The data are presented as the mean ± SEM. Every microwell condition was compared to control using Student's t-test. *P < 0.05; **P < 0.01; ***P < 0.001; ns, no significance. Scale bars (A: 100 μm, E: 50 μm). Note, all data were acquired with at least 3 organoids for each condition.