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KMID : 0877920120140040192
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Korean Journal of Ultrasound in Obstetrics and Gynocology
2012 Volume.14 No. 4 p.192 ~ p.199
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A Novel Reproductive Toxicology Test Using Amnionic Epithelial Stem Cells (AESCs) for Predicting Teratogenic Effect Of Medications
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Chung Ye-Won
Han Song-Yee
Cho Geum-Joon
Hong Soon-Cheol
Han Jeong-Yeol
Oh Min-Jeong
Kim Hai-Joong
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Abstract
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Objective: One of the most important purposes of antenatal screening is to detect fetal anomaly. The classic screening test for detecting such anomaly is ultrasound until now. And there are some medical conditions such as epilepsy that can’t be avoid medication during pregnancy. But the best screening strategy for predicting teratogenicity of various medications is not established yet. In this study, we performed MTT assay using human amnionic epithelial stem cells (AESCs) to assess the cytotoxicity of the various antiepileptic drug and to verify the availability of reproductive toxicity test using AESCs.
Methods: AESCs were isolated and cultured from the human amnion. To examine phenotype of the AESCs, characteristics of the cultured AESCs were identified using FACS analysis. Neuronal and adipogenic differentiation of AESCs were accomplished under appropriate culture conditions. After characterization of AESCs, cytotoxicity of Lamotrigine was evaluated using MTT assay with AESCs, compared to those of Valproic acid and Topiramate.
Results: After isolation of AESCs, the characteristics were evaluated using FACS analysis. The AESCs were positive for pluripotent stem cell markers, Oct-4, Nanog, SSEA-4 and positive for CD73, CD90, CD105, CD324, SOX-2. While negative for CD349 that mean the epithelial origin’s stem cells. The neurogenic differentiation were achieved and it were confirmed by immunocytochemistry staining using antibodies to MAP2 and GFAP, which is the marker for neuron and neurogliocyte, respectively. In addition, adipogenic differentiation was also achieved with confirming Oil red-O staining. Cell cytotoxicity test using MTT assay showed that IC50 of lamotrigine is 1 mM, which indicated relatively stronger cytotoxicity comparing with IC50 of other anticonvulsants, valproic acid (10 mM) and topiramate (5 mM).
Conclusion: We could confirm the pluripotency of AESCs. And to expect embryonic toxicity of new medication, AESCs is one of the most useful source of the stem cells which could be applied to human reproductive toxicity test.
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KEYWORD
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Fetal anomaly, Reproductive toxicity test, Amnionic epithelial stem cells
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