The Effect Of Retinoic Acid Human Embryonic Stem Cells With And Without Rosette Morphology

Document Type : Original Article

Authors

Royan Institute, Stem Cell Department, Tehran, Iran

Abstract

Introduction: Human Embryonic Stem (hES) Cells are pluripotent cells derived form the inner cell mass of blastocyst. These unique cells have the potential to form virtually any cell type in the body and can be propagated in vitro indefinitely in an undifferentiated state. This study was initiated to assess the effect of retinoic acid (RA) on hES cells with and without rosette morphology (neuroectodermal – like cells).
Material and Methods: Embryoid bodies (EBs) were made from ES cell line (Royan H1) with and without rosset compartments. The EBs treated with retinoic acid (RA) (10 µM) for 20 days and the cultured on tissue culture plate individually containing neuronal medium. Differentiated cells were evaluated by phase contrast microscopy and immunocytochemistry.
Results: Differentiated cells were positive with the neuron specific antibodies against microtubules associated protein 2 (a + b) (MAP – 2) , neurofilament protein (NP), neuron – specific enolase (NSE), synaptophysin, and tubulin III. In presence of RA, more EBs were differentiated into neurons in both groups (in group with rosette compartment, 73.2% v.s. 13.2%, P<0.01, and in group with non – rosette compartment, 57.5% v.s. 19.5% P< 0.01). Moreover, RA in EBs with rosette compartment increased neural morphology.
Conclusion: These results showed RA induces hES into neurogenesis and rosette compartments f hES have more potential for neurogenesis and these cells may be used in stem cell transplantation therapies for diseases in future.

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