Development and Maintenance of Serum-Free Embryoid Bodies
Transcription
Begin with a well containing neural progenitor cell or NPC aggregates in differentiating media-1 or DM-1.
These NPCs are precursors of neural lineage generated from human induced pluripotent stem cells or hiPSCs using DM-1 under controlled conditions.
Transfer the NPC aggregates onto a membrane insert placed inside a well containing DM-1 and remove excess media.
Change the medium to DM-2 and incubate.
DM-2 provides nutrients and differentiation factors that promote the maturation of NPCs into different neural cell types.
Undifferentiated NPCs transform into embryoid bodies, known as serum-free embryoid bodies or SFEBs, as the media lacks serum. These EBs resemble the morphology of a developing brain.
Transfer the insert into a well containing DM-3.
DM-3 contains growth factors essential for the maintenance and growth of SFEBs.
Incubate till SFEBs reach the desired size.
