Stem Cells and Development
P53 Mutation and Epigenetic Imprinted IGF2/H19 Gene Analysis in Mesenchymal Stem Cells Derived from Amniotic Fluid, Amnion, Endometrium, and Wharton's Jelly
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Phermthai Tatsanee, Pokathikorn Puttachart, Wichitwiengrat Suparat, Thongbopit Sasiprapa, Tungprasertpol Kittima, and Julavijitphong Suphakde. Stem Cells and Development. June 2017, ahead of print.http://ift.tt/2ueH2c2
Phermthai Tatsanee, Pokathikorn Puttachart, Wichitwiengrat Suparat, Thongbopit Sasiprapa, Tungprasertpol Kittima, and Julavijitphong Suphakde. Stem Cells and Development. June 2017, ahead of print.http://ift.tt/2ueH2c2
Online Ahead of Editing: June 19, 2017
Online Ahead of Print: June 14, 2017
Online Ahead of Print: June 14, 2017
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Tatsanee Phermthai, Puttachart Pokathikorn, Suparat Wichitwiengrat, Sasiprapa Thongbopit, Kittima Tungprasertpol, and Suphakde Julavijitphong
Stem Cell Research and Development Unit, Obstetrics and Gynecology Department, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, Thailand.
Received for publication January 29, 2017
Accepted after revision May 30, 2017
Accepted after revision May 30, 2017
ABSTRACT
Mesenchymal stem cells (MSC) are promising cells for medical therapy. In in vitro expansion, MSC can give rise to progeny with genomic and epigenomic alterations, resulting in senescence, loss of terminal differentiation, and transformation to cancer. However, MSC genome protects its genetic instability by a guardian function of the P53tumor suppressor gene and epigenetic balance system during MSC culture. Mutations of P53 and epigenetic alterations have been reported to disrupt the quality and quantity of MSC and initiate tumorigenesis. We monitor P53 and epigenetic changes in MSC derived from amniotic fluid (AF-MSC), amnion membrane (AM-MSC), endometrium (EM-MSC), and Wharton's jelly (WJ-MSC) by the missense mutation analysis of the P53 gene and the expression levels of P53, and epigenetic insulin-like growth factor 2 (IGF2) and H19-imprinted genes. Our work demonstrates a variation of P53 expression among different MSC types. AF-MSC has a high P53 expression level with retaining a stability of P53expression throughout a long culture period, whereas EM-MSC and WJ-MSC showed variation of P53 gene expression during culture. Epigenetic analysis showed a stable H19 expression pattern in AF-MSC, AM-MSC, and EM-MSC culture, whereas H19 expression fluctuated in WJ-MSC culture. We conclude that gene instability can be found during in vitro MSC expansion. With awareness to MSC quality and safety in MSC transformation risk, P53 mutation and IGF2 and H19-imprinted gene analysis should be applied to monitor in therapeutic-grade MSC. We also demonstrated that AF-MSC is one of the most interesting MSC for medical therapy because of its high genomic stability and epigenetic fidelity.
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