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. 2015 Nov 12;6(2):228–242. doi: 10.1016/j.stemcr.2015.10.007

Figure 2.

Figure 2

Single NESCs Self-Organize into Miniature NT-like Structures

Three independent experiments were repeated for NESCs from IVF3.2 and IVF3.3.

(A) Schematic representation of single-NESC self-organization into NT structures in the continual colony assays.

(B) A NESC on one well of 96-well plate at day 1.

(C and D) Representative one-cell-derived NT structure at day 14.

(E–H) Single-cell-derived NTs expressed NESC markers PAX6 (E), NESTIN (F), N-CADHERIN (G), and ZO-1 (H).

(I) BrdU incorporation of single-cell-derived NTs.

(J) Live imaging of NESC interkinetic nuclear migration in an NT (also see Movie S2). NESC division predominantly displayed horizontal (close to apical surface) orientation. Red arrows indicate two dividing cells; red lines indicate dividing axis; yellow arrows indicate two divided cells; yellow scales indicate the distance between a lumen and GFP+ cells.

(K) Percentages of single-seeded cells on day 14 that formed NESC-derived survival colonies and NT colonies at sequential passages.

(L) Percentage of inoculated single cells that formed survival colonies (phase 1) and the percentage of survival colonies that formed NT colonies (phase 2) during the continual colony assays.

Data are shown as mean ± SD (data from three independent experiments). No significant difference was found (p > 0.05). Scale bars: (C and I), 100 μm; others, 20 μm.