Figure 4.

Longitudinal Assessment of hiPSC Morphogenesis Reveals Hypertrophy in Cell-Cycle Growth Phase, Confluency-Induced Shape Changes, and Colony Migration

(A) Cell area measurements spanning two cell divisions with cell area measured every 12 min. The duration of each cells' cell-cycle length was normalized to the percentage of the cell cycle. Gray lines indicate individual cells, black line indicates line of best fit, and dashed vertical lines indicate cytokinesis time points at 0% and 100% of the cell cycle.

(B) Cell area measurements over 8 days in culture.

(C) Examples of 3D reconstructions at days 0 and 7 after plating (601 × 601 μm, 0.4-μm steps). Scale bar, 20 μm.

(D) Quantification of cell height over 8 days.

(E and F) Migration analysis tracking absolute (E) or relative (F) positions of individual hiPSCs.

(G) Representative image showing an example of one parental cell on the colony perimeter at 0 h that gave rise to two internally localized cells at 12 h (white arrows) and another that gave rise to two cells with maintained peripheral localization (white arrowheads). Scale bars, 20 μm.

(H) Analysis of edge PSCs showing the proportion remaining on the edge or translocating interiorly at each time point.

(I and J) Migration analysis tracking absolute (I) or relative (J) hiPSC colony centroid position from days 0 to 4. (I) “X” denotes colony start position and arrowheads indicate colony end position. Colony fusion events are indicated by dashed lines and split color arrowheads.