Proliferation and survival of human amniotic epithelial cells during their hepatic differentiation

Stem cells derived from placental tissues are an attractive source of cells for regenerative medicine. Amniotic epithelial cells isolated from human amnion (hAECs) have desirable and competitive characteristics that make them stand out between other stem cells. They have the ability to differentiate...

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Autores principales:	Maymó, J.L., Riedel, R., Pérez-Pérez, A., Magatti, M., Maskin, B., Dueñas, J.L., Parolini, O., Sánchez-Margalet, V., Varone, C.L.
Formato:	JOUR
Materias:	cyclin D1 Ki 67 antigen mitogen activated protein kinase 1 mitogen activated protein kinase 3 octamer transcription factor 4 protein p21 protein p53 stage specific embryo antigen 4 transcription factor NANOG transcription factor Sox2 biological marker amnion cell amniotic epithelial cell Article cell culture cell cycle cell differentiation cell proliferation cell structure cell survival controlled study enzyme phosphorylation human human cell immunofluorescence liver cell pluripotent stem cell protein expression reverse transcription polymerase chain reaction upregulation Western blotting amnion cytology epithelium cell female liver MAPK signaling metabolism phosphorylation pregnancy real time polymerase chain reaction Amnion Biomarkers Cell Proliferation Cell Survival Cells, Cultured Epithelial Cells Female Humans Liver MAP Kinase Signaling System Phosphorylation Pregnancy Real-Time Polymerase Chain Reaction
Acceso en línea:	http://hdl.handle.net/20.500.12110/paper_19326203_v13_n1_p_Maymo
Aporte de:	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) de Universidad de Buenos Aires

id	todo:paper_19326203_v13_n1_p_Maymo
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spelling	todo:paper_19326203_v13_n1_p_Maymo2023-10-03T16:34:55Z Proliferation and survival of human amniotic epithelial cells during their hepatic differentiation Maymó, J.L. Riedel, R. Pérez-Pérez, A. Magatti, M. Maskin, B. Dueñas, J.L. Parolini, O. Sánchez-Margalet, V. Varone, C.L. cyclin D1 Ki 67 antigen mitogen activated protein kinase 1 mitogen activated protein kinase 3 octamer transcription factor 4 protein p21 protein p53 stage specific embryo antigen 4 transcription factor NANOG transcription factor Sox2 biological marker amnion cell amniotic epithelial cell Article cell culture cell cycle cell differentiation cell proliferation cell structure cell survival controlled study enzyme phosphorylation human human cell immunofluorescence liver cell pluripotent stem cell protein expression reverse transcription polymerase chain reaction upregulation Western blotting amnion cytology epithelium cell female liver MAPK signaling metabolism phosphorylation pregnancy real time polymerase chain reaction Amnion Biomarkers Cell Proliferation Cell Survival Cells, Cultured Epithelial Cells Female Humans Liver MAP Kinase Signaling System Phosphorylation Pregnancy Real-Time Polymerase Chain Reaction Stem cells derived from placental tissues are an attractive source of cells for regenerative medicine. Amniotic epithelial cells isolated from human amnion (hAECs) have desirable and competitive characteristics that make them stand out between other stem cells. They have the ability to differentiate toward all three germ layers, they are not tumorigenic and they have immunosuppressive properties. Although liver transplantation is the best way to treat acute and chronic hepatic failure patients, there are several obstacles. Recently, stem cells have been spotlighted as alternative source of hepatocytes because of their potential for hepatogenic differentiation. In this work, we aimed to study the proliferation and survival of the hAECs during their hepatic differentiation. We have also analyzed the changes in pluripotency and hepatic markers. We differentiated amniotic cells applying a specific hepatic differentiation (HD) protocol. We determined by qRT-PCR that hAECs express significant levels of SOX-2, OCT-4 and NANOG during at least 15 days in culture and these pluripotent markers diminish during HD. SSEA-4 expression was reduced during HD, measured by immunofluorescence. Morphological characteristics became more similar to hepatic ones in differentiated cells and representative hepatic markers significantly augmented their expression, measured by qRT-PCR and Western blot. Cells achieved a differentiation efficiency of 75%. We observed that HD induced proliferation and promoted survival of hAECs, during 30 days in culture, evaluated by 3H-thymidine incorporation and MTT assay. HD also promoted changes in hAECs cell cycle. Cyclin D1 expression increased, while p21 and p53 levels were reduced. Immunofluorescence analysis showed that Ki-67 expression was upregulated during HD. Finally, ERK 1/2 phosphorylation, which is intimately linked to proliferation and cell survival, augmented during all HD process and the inhibition of this signaling pathway affected not only proliferation but also differentiation. Our results suggest that HD promotes proliferation and survival of hAECs, providing important evidence about the mechanisms governing their hepatic differentiation. We bring new knowledge concerning some of the optimal transplantation conditions for these hepatic like cells. Copyright: © 2018 Maymó et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. JOUR info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by/2.5/ar http://hdl.handle.net/20.500.12110/paper_19326203_v13_n1_p_Maymo
institution	Universidad de Buenos Aires
institution_str	I-28
repository_str	R-134
collection	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA)
topic	cyclin D1 Ki 67 antigen mitogen activated protein kinase 1 mitogen activated protein kinase 3 octamer transcription factor 4 protein p21 protein p53 stage specific embryo antigen 4 transcription factor NANOG transcription factor Sox2 biological marker amnion cell amniotic epithelial cell Article cell culture cell cycle cell differentiation cell proliferation cell structure cell survival controlled study enzyme phosphorylation human human cell immunofluorescence liver cell pluripotent stem cell protein expression reverse transcription polymerase chain reaction upregulation Western blotting amnion cytology epithelium cell female liver MAPK signaling metabolism phosphorylation pregnancy real time polymerase chain reaction Amnion Biomarkers Cell Proliferation Cell Survival Cells, Cultured Epithelial Cells Female Humans Liver MAP Kinase Signaling System Phosphorylation Pregnancy Real-Time Polymerase Chain Reaction
spellingShingle	cyclin D1 Ki 67 antigen mitogen activated protein kinase 1 mitogen activated protein kinase 3 octamer transcription factor 4 protein p21 protein p53 stage specific embryo antigen 4 transcription factor NANOG transcription factor Sox2 biological marker amnion cell amniotic epithelial cell Article cell culture cell cycle cell differentiation cell proliferation cell structure cell survival controlled study enzyme phosphorylation human human cell immunofluorescence liver cell pluripotent stem cell protein expression reverse transcription polymerase chain reaction upregulation Western blotting amnion cytology epithelium cell female liver MAPK signaling metabolism phosphorylation pregnancy real time polymerase chain reaction Amnion Biomarkers Cell Proliferation Cell Survival Cells, Cultured Epithelial Cells Female Humans Liver MAP Kinase Signaling System Phosphorylation Pregnancy Real-Time Polymerase Chain Reaction Maymó, J.L. Riedel, R. Pérez-Pérez, A. Magatti, M. Maskin, B. Dueñas, J.L. Parolini, O. Sánchez-Margalet, V. Varone, C.L. Proliferation and survival of human amniotic epithelial cells during their hepatic differentiation
topic_facet	cyclin D1 Ki 67 antigen mitogen activated protein kinase 1 mitogen activated protein kinase 3 octamer transcription factor 4 protein p21 protein p53 stage specific embryo antigen 4 transcription factor NANOG transcription factor Sox2 biological marker amnion cell amniotic epithelial cell Article cell culture cell cycle cell differentiation cell proliferation cell structure cell survival controlled study enzyme phosphorylation human human cell immunofluorescence liver cell pluripotent stem cell protein expression reverse transcription polymerase chain reaction upregulation Western blotting amnion cytology epithelium cell female liver MAPK signaling metabolism phosphorylation pregnancy real time polymerase chain reaction Amnion Biomarkers Cell Proliferation Cell Survival Cells, Cultured Epithelial Cells Female Humans Liver MAP Kinase Signaling System Phosphorylation Pregnancy Real-Time Polymerase Chain Reaction
description	Stem cells derived from placental tissues are an attractive source of cells for regenerative medicine. Amniotic epithelial cells isolated from human amnion (hAECs) have desirable and competitive characteristics that make them stand out between other stem cells. They have the ability to differentiate toward all three germ layers, they are not tumorigenic and they have immunosuppressive properties. Although liver transplantation is the best way to treat acute and chronic hepatic failure patients, there are several obstacles. Recently, stem cells have been spotlighted as alternative source of hepatocytes because of their potential for hepatogenic differentiation. In this work, we aimed to study the proliferation and survival of the hAECs during their hepatic differentiation. We have also analyzed the changes in pluripotency and hepatic markers. We differentiated amniotic cells applying a specific hepatic differentiation (HD) protocol. We determined by qRT-PCR that hAECs express significant levels of SOX-2, OCT-4 and NANOG during at least 15 days in culture and these pluripotent markers diminish during HD. SSEA-4 expression was reduced during HD, measured by immunofluorescence. Morphological characteristics became more similar to hepatic ones in differentiated cells and representative hepatic markers significantly augmented their expression, measured by qRT-PCR and Western blot. Cells achieved a differentiation efficiency of 75%. We observed that HD induced proliferation and promoted survival of hAECs, during 30 days in culture, evaluated by 3H-thymidine incorporation and MTT assay. HD also promoted changes in hAECs cell cycle. Cyclin D1 expression increased, while p21 and p53 levels were reduced. Immunofluorescence analysis showed that Ki-67 expression was upregulated during HD. Finally, ERK 1/2 phosphorylation, which is intimately linked to proliferation and cell survival, augmented during all HD process and the inhibition of this signaling pathway affected not only proliferation but also differentiation. Our results suggest that HD promotes proliferation and survival of hAECs, providing important evidence about the mechanisms governing their hepatic differentiation. We bring new knowledge concerning some of the optimal transplantation conditions for these hepatic like cells. Copyright: © 2018 Maymó et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
format	JOUR
author	Maymó, J.L. Riedel, R. Pérez-Pérez, A. Magatti, M. Maskin, B. Dueñas, J.L. Parolini, O. Sánchez-Margalet, V. Varone, C.L.
author_facet	Maymó, J.L. Riedel, R. Pérez-Pérez, A. Magatti, M. Maskin, B. Dueñas, J.L. Parolini, O. Sánchez-Margalet, V. Varone, C.L.
author_sort	Maymó, J.L.
title	Proliferation and survival of human amniotic epithelial cells during their hepatic differentiation
title_short	Proliferation and survival of human amniotic epithelial cells during their hepatic differentiation
title_full	Proliferation and survival of human amniotic epithelial cells during their hepatic differentiation
title_fullStr	Proliferation and survival of human amniotic epithelial cells during their hepatic differentiation
title_full_unstemmed	Proliferation and survival of human amniotic epithelial cells during their hepatic differentiation
title_sort	proliferation and survival of human amniotic epithelial cells during their hepatic differentiation
url	http://hdl.handle.net/20.500.12110/paper_19326203_v13_n1_p_Maymo
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_version_	1782027152116416512

Proliferation and survival of human amniotic epithelial cells during their hepatic differentiation

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