Gene Expression Patterns of Royan Human Embryonic Stem Cells Correlate with Their Propensity and Culture Systems

Objective Human embryonic stem cells (hESCs) have the potential to give rise to all types of cells in the human body when appropriately induced to differentiate. Stem cells can differentiate spontaneously into the threegerm layer derivatives by embryoid bodies (EBs) formation. However, the twodimensional (2D) adherent culture of hESCs under defined conditions is commonly used for directed differentiation toward a specific type of mature cells. In this study, we aimed to determine the propensity of the Royan hESC lines based on comparison of expression levels of 46 lineage specific markers. Materials and Methods In this experimental study, we have compared the expression of lineagespecific markers in hESC lines during EB versus adherentbased spontaneous differentiation. We used quantitative realtime polymerase chain reaction (qRTPCR) to assess expressions of 46 lineagespecific markers in 4 hESC lines, Royan H1 (RH1), RH2, RH5, and RH6, during spontaneous differentiation in both EB and adherent cultures at 0, 10, and 30 days after initiation of differentiation. Results Based on qRTPCR data analysis, the liver and neuronal markers had higher expression levels in EBs, whereas skinspecific markers expressed at higher levels in the adherent culture. The results showed differential expression patterns of some lineagespecific markers in EBs compared with the adherent cultures. ConclusionAccording to these results, possibly the spontaneous differentiation technique could be a useful method for optimization of culture conditions to differentiate stem cells into specific cell types such ectoderm, neuron, endoderm and hepatocyte. This approach might prove beneficial for further work on maximizing the efficiency of directed differentiation and development of novel differentiation protocols.