The protective effect of folic acid in human amniotic epithelial stem cells using a new reproductive toxicity test

다국어 초록 (Multilingual Abstract)

The amnion arises from the inner cell mass of blastocyst; therefore, its embryological origin is similar to that of the human foetus, and it has similar potential to differentiate into all three germ layers. Human-derived amniotic epithelial stem cells (hAESCs) are rich in amnion, but unlike embryonic stem cells, they are not associated with the ethical issues that have impeded stem cell research. We performed lamotrigine cytotoxicity tests with AESCs and developmental neurotoxicity tests during spontaneous differentiation of hAESCs to neuronal cells in order to measure cytotoxicity and confirm the protective effect of folic acid. The hAESCs isolated from amnion tissue were characterized by flow cytometry. Lamotrigine cytotoxicity was measured at concentrations of 10 μM, 50 μM, 100 μM, 500 μM, and 1000 μM; folic acid was added at 10 nM, 100 nM, and 1000 nM and cell viability was measured 1, 2, and 3 days after the treatment. MTT assays were used to measure the protective effect of folic acid in cells treated with lamotrigine. During neuronal differentiation of AESCs, lamotrigine and folic acid were added and the cells were evaluated for neurotoxicity and neuroprotection by folic acid. The hAESCs were positive for CD73, CD90, CD105, CD324, SSEA-4, SOX-2, OCT-3/4, and Nanog; they were negative for CD19, CD34, and CD349. The cytotoxicity assay showed that cell survival significantly reduced at 500μM and 1000 μM lamotrigine. Pre-treatment with 10–100 nM folic acid showed maximal protective effect. With 500 μM lamotrigine, neuronal differentiation reduced by ~30%. Early supplementation with folic acid (10 nM) did not improve neuronal differentiation. These results suggest that hAESCs could be used to evaluate reproductive toxicity in vitro.

목차 (Table of Contents)

• Abstract ------------------------------------------------i
• Introduction -------------------------------------------1
• Materials and methods -------------------------------2
• Results -------------------------------------------------4
• Discussion ---------------------------------------------6
• References --------------------------------------------12