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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
  1. Protocol illustrated in Fig. 1. Culture medium was changed daily until pellet formation. Pellet medium was changed every other day
  2. BMP bone morphogenetic protein 4, iPSC induced pluripotent stem cell, FGF2 basic fibroblast growth factor, NP nucleus pulposus, TGF transforming growth factor