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

چکیده مقاله

Abstract
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 three-germ layer
derivatives by embryoid bodies (EBs) formation. However, the two-dimensional (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 lineage-specific markers in
hESC lines during EB versus adherent-based spontaneous differentiation. We used quantitative real-time polymerase
chain reaction (qRT-PCR) to assess expressions of 46 lineage-specific 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 qRT-PCR data analysis, the liver and neuronal markers had higher expression levels in EBs,
whereas skin-specific markers expressed at higher levels in the adherent culture. The results showed differential
expression patterns of some lineage-specific markers in EBs compared with the adherent cultures.
Conclusion: According 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.