FIG. 1.

Generation and aggregation of neural progenitor cells (NPCs) and V2a interneurons (INs). (A) NPCs isolated from E13.5 rat spinal cords were stored frozen and thawed 1 day prior to aggregation. (B) Chx10-Puro embryonic stem cells (ESCs) expressing TdTomato were induced using a 2^-/4⁺ protocol followed by 24 h of puromycin selection for V2a INs. (C) Schematic showing aggregation protocol. Populations of NPCs were mixed at a 1:1 ratio with TdTomato-V2a cells and seeded into an AggreWell plate. After 2 days of culture, spherical aggregates form in the dish (green fluorescent protein/TdTomato, NPC/V2a INs, respectively shown in D-F), which are then resuspended in fresh media and washed in Hank's Balanced Salt Solution prior to transplantation (F). Scale bars are as indicated. (G) Experimental timeline of the study.