hESC-specific phenotypic markers, such as Oct4 and SSEA4, were evaluated qualitatively by indirect immunofluorescence staining (Figure 2A,B) and quantitatively by flow cytometry (Figure 2C,D). After ten passages on Synthemax Surface in X-VIVO10+GF medium, H7 hESCs retained high levels of both markers, suggesting that they maintained their undifferentiated status.
These results demonstrate that H7 hESC can be successfully propagated on Synthemax Surface for multiple passages in xeno-free, defined medium, while retaining important hESC characteristics, such as stable proliferation rate, phenotypic marker profile, and normal karyotype. Similar results were demonstrated with other hESC lines (H1, H9, BG01v) and defined media conditions (mTeSR®1, STEMPRO® hESC SFM, and NutriStem™).
Pluripotency, the ability to differentiate into cells of all three germ layers, is a fundamental property of hESCs. This property is a critical parameter when evaluating new culture conditions for hESCs.