Human stem cells cultured in NM23-H1-MM over anti-MUC1* antibody surfaces express higher levels of naïve markers and lower levels of primed markers.

RT-PCR was used to quantify expression of a subset of naïve markers that included Oct4, Nanog, Klf4 and Klf2, which should be high in the naïve state, and a subset of primed markers that included FoxA2, XIST, Otx2 and Lhx2, which are high in the primed state but low in the naïve state. Measurements were normalized to housekeeping gene GAPDH and expressed as fold change to H9 ES cells cultured in 4ng/ml bFGF over MEFs (control, n = 3). a) H9 ES cells cultured in NM23-H1-MM on anti-MUC1* antibody (MN-C3) surfaces, on average, showed increased expression of naïve markers and decreased expression of primed markers (n = 6). Conversely, H9 cells cultured in mTeSR over Matrigel showed decreased expression of naïve markers and increased expression of primed markers (n = 4). b) Individual measurements of the subset of naïve or primed markers are plotted as a function of passage number for NM23-H1-MM over anti-MUC1* antibody surfaces and c) for mTeSR over Matrigel. The trend toward the naïve state increased with successive passage in NM23-H1-MM but not with mTeSR. Large standard error for some experiments may be due to contamination of visually pluripotent stem cells with newly differentiating stem cells. For statistical analysis see Fig. S10 and S11.