[Induced in vitro differentiation of beta -tubulin III expressed cells from human amniotic epithelial cells]

ABSTRACT

The differentiation of neural cells from embryonic stem cells is influenced by several factors. Amniotic epithelial cells [AECs] share many of the same characteristics as embryonic stem cells, and therefore, those factors may similarly affect the derivation of neural cells from AECs. In this study, we examined the differentiation of neural cells in vitro from AECs using the expression of Beta-tubulin III marker, after AECs treatment with retinoic acid [RA], and the impact of basic fibroblast growth factor [bFGF]. We also studied whether blocking bone morphogenetic protein [BMP] signaling the use of its antagonist, noggin, affects the derivation of neural cells from AECs. AECs were isolated from fresh human amniotic membrane and aggregated for 5 days in bacteriological dishes. The dissociated cells were transferred to adherent culture dishes and treated with noggin and bFGF for 7 days. In some cultures, bFGF was removed and RA was added and the cultures were allowed to differentiate for 21 days. Analysis of AECs derived neural cells was performed at the Beta-tubulin III expression levels by immunocitochemistry. All cultures treated with noggin showed the higher levels of Beta-tubulin III expression than noggin free cultures. Combined treatment with bFGF and RA showed the highest level of Beta-tubulin III in all treatment groups with or without noggin. bFGF withdrawal did not promote expression of Beta-tubulin III, while its maintenance increased the expression of Beta-tubulin III. These results show the capability of AECs to express neural cell marker and this potential is affected by some factors including noggin, bFGF, RA