Table 1 Nucleus pulposus differentiation media defined by factors added to cell culture to promote differentiation of iPSCs into NP-like cells in a directed, stepwise differentiation protocol as illustrated in Fig. 1. Culture medium was changed daily until pellet formation. Pellet medium was changed every other day
From: Differentiation of human induced pluripotent stem cells into nucleus pulposus-like cells
Stage | Day | Wnt-3a (ng/ml) | ActivinA (ng/ml) | FGF2 (ng/ml) | BMP4 (ng/ml) | Follistatin (ng/ml) | GDF5 (ng/ml) | TGF-β3 (ng/ml) |
|---|---|---|---|---|---|---|---|---|
Step 1a: notochord/mesoderm differentiation | 1 | 25 | 50 | – | – | – | – | – |
2 | 25 | 25 | 20 | 40 | – | – | – | |
3 | 25 | 10 | 20 | 40 | – | – | – | |
Step 1b: notochord/mesoderm differentiation | 4 | – | 5 | 20 | 40 | 100 | – | – |
5 | – | 5 | 20 | 40 | 100 | – | – | |
6 | – | 2 | 20 | 40 | 100 | – | – | |
7 | – | 2 | 20 | 40 | 100 | – | – | |
Step 2: expansion in monolayer and differentiation | Replate 8 | – | 2 | 20 | 20 | – | – | – |
9–14 | – | – | 5 | – | – | 40 | – | |
Step 3: pellet culture and differentiation | 15–36 | – | – | – | – | – | – | 10 |