Robust Differentiation in Chemically Defined Conditions across Multiple hPSC Lines
(A and B) CM content (A) and respective cell numbers (B) on day 10 using bioreactor-derived aggregates from day-2 precultures of three cell lines differentiated in Erlenmeyer flasks in two or three independent experiments per cell line. (A) hESCs p = 0.1972, Phoenix cells p = 0.0575. (B) hESCs p = 0.8835, Pheonix cells p = 0.0067.
(C) Robustness of the chemically defined differentiation protocol was confirmed by FC for three markers with >70% CM found for seven cell lines tested in agitated 6-well plates in 2–5 independent experiments: hES3, bMHC_iPS, SFC840: n = 2; H9, CBiPS2, HDFiPS: n = 3; Phoenix: n = 5.
(D) Comparison of IWP2 versus Wnt-C59 added after 24 h of 5 μM CHIR treatment in three independent experiments revealed similar efficiencies in 6-well plates on day 10 for Phoenix.
Error bars represent SEM.