Markers of Pluripotency in Human Amniotic Epithelial Cells and Their Differentiation to Progenitor of Cortical Neurons.
Human pluripotent stem cells (hPSC) have promise for regenerative medicine due to their auto-renovation and differentiation capacities. Nevertheless, there are several ethical and methodological issues about these cells that have not been resolved. Human amniotic epithelial cells (hAEC) have been pr...
| Main Authors: | , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Public Library of Science (PLoS)
2015-01-01
|
| Series: | PLoS ONE |
| Online Access: | http://europepmc.org/articles/PMC4697857?pdf=render |
| _version_ | 1818950728436154368 |
|---|---|
| author | Irma Lydia García-Castro Guadalupe García-López Daniela Ávila-González Héctor Flores-Herrera Anayansi Molina-Hernández Wendy Portillo Eva Ramón-Gallegos Néstor Fabián Díaz |
| author_facet | Irma Lydia García-Castro Guadalupe García-López Daniela Ávila-González Héctor Flores-Herrera Anayansi Molina-Hernández Wendy Portillo Eva Ramón-Gallegos Néstor Fabián Díaz |
| author_sort | Irma Lydia García-Castro |
| collection | DOAJ |
| description | Human pluripotent stem cells (hPSC) have promise for regenerative medicine due to their auto-renovation and differentiation capacities. Nevertheless, there are several ethical and methodological issues about these cells that have not been resolved. Human amniotic epithelial cells (hAEC) have been proposed as source of pluripotent stem cells. Several groups have studied hAEC but have reported inconsistencies about their pluripotency properties. The aim of the present study was the in vitro characterization of hAEC collected from a Mexican population in order to identify transcription factors involved in the pluripotency circuitry and to determine their epigenetic state. Finally, we evaluated if these cells differentiate to cortical progenitors. We analyzed qualitatively and quantitatively the expression of the transcription factors of pluripotency (OCT4, SOX2, NANOG, KLF4 and REX1) by RT-PCR and RT-qPCR in hAEC. Also, we determined the presence of OCT4, SOX2, NANOG, SSEA3, SSEA4, TRA-1-60, E-cadherin, KLF4, TFE3 as well as the proliferation and epigenetic state by immunocytochemistry of the cells. Finally, hAEC were differentiated towards cortical progenitors using a protocol of two stages. Here we show that hAEC, obtained from a Mexican population and cultured in vitro (P0-P3), maintained the expression of several markers strongly involved in pluripotency maintenance (OCT4, SOX2, NANOG, TFE3, KLF4, SSEA3, SSEA4, TRA-1-60 and E-cadherin). Finally, when hAEC were treated with growth factors and small molecules, they expressed markers characteristic of cortical progenitors (TBR2, OTX2, NeuN and β-III-tubulin). Our results demonstrated that hAEC express naïve pluripotent markers (KLF4, REX1 and TFE3) as well as the cortical neuron phenotype after differentiation. This highlights the need for further investigation of hAEC as a possible source of hPSC. |
| first_indexed | 2024-12-20T09:23:12Z |
| format | Article |
| id | doaj.art-7908d4bd400540088fd5a7d716d7b2cb |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-12-20T09:23:12Z |
| publishDate | 2015-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-7908d4bd400540088fd5a7d716d7b2cb2022-12-21T19:45:14ZengPublic Library of Science (PLoS)PLoS ONE1932-62032015-01-011012e014608210.1371/journal.pone.0146082Markers of Pluripotency in Human Amniotic Epithelial Cells and Their Differentiation to Progenitor of Cortical Neurons.Irma Lydia García-CastroGuadalupe García-LópezDaniela Ávila-GonzálezHéctor Flores-HerreraAnayansi Molina-HernándezWendy PortilloEva Ramón-GallegosNéstor Fabián DíazHuman pluripotent stem cells (hPSC) have promise for regenerative medicine due to their auto-renovation and differentiation capacities. Nevertheless, there are several ethical and methodological issues about these cells that have not been resolved. Human amniotic epithelial cells (hAEC) have been proposed as source of pluripotent stem cells. Several groups have studied hAEC but have reported inconsistencies about their pluripotency properties. The aim of the present study was the in vitro characterization of hAEC collected from a Mexican population in order to identify transcription factors involved in the pluripotency circuitry and to determine their epigenetic state. Finally, we evaluated if these cells differentiate to cortical progenitors. We analyzed qualitatively and quantitatively the expression of the transcription factors of pluripotency (OCT4, SOX2, NANOG, KLF4 and REX1) by RT-PCR and RT-qPCR in hAEC. Also, we determined the presence of OCT4, SOX2, NANOG, SSEA3, SSEA4, TRA-1-60, E-cadherin, KLF4, TFE3 as well as the proliferation and epigenetic state by immunocytochemistry of the cells. Finally, hAEC were differentiated towards cortical progenitors using a protocol of two stages. Here we show that hAEC, obtained from a Mexican population and cultured in vitro (P0-P3), maintained the expression of several markers strongly involved in pluripotency maintenance (OCT4, SOX2, NANOG, TFE3, KLF4, SSEA3, SSEA4, TRA-1-60 and E-cadherin). Finally, when hAEC were treated with growth factors and small molecules, they expressed markers characteristic of cortical progenitors (TBR2, OTX2, NeuN and β-III-tubulin). Our results demonstrated that hAEC express naïve pluripotent markers (KLF4, REX1 and TFE3) as well as the cortical neuron phenotype after differentiation. This highlights the need for further investigation of hAEC as a possible source of hPSC.http://europepmc.org/articles/PMC4697857?pdf=render |
| spellingShingle | Irma Lydia García-Castro Guadalupe García-López Daniela Ávila-González Héctor Flores-Herrera Anayansi Molina-Hernández Wendy Portillo Eva Ramón-Gallegos Néstor Fabián Díaz Markers of Pluripotency in Human Amniotic Epithelial Cells and Their Differentiation to Progenitor of Cortical Neurons. PLoS ONE |
| title | Markers of Pluripotency in Human Amniotic Epithelial Cells and Their Differentiation to Progenitor of Cortical Neurons. |
| title_full | Markers of Pluripotency in Human Amniotic Epithelial Cells and Their Differentiation to Progenitor of Cortical Neurons. |
| title_fullStr | Markers of Pluripotency in Human Amniotic Epithelial Cells and Their Differentiation to Progenitor of Cortical Neurons. |
| title_full_unstemmed | Markers of Pluripotency in Human Amniotic Epithelial Cells and Their Differentiation to Progenitor of Cortical Neurons. |
| title_short | Markers of Pluripotency in Human Amniotic Epithelial Cells and Their Differentiation to Progenitor of Cortical Neurons. |
| title_sort | markers of pluripotency in human amniotic epithelial cells and their differentiation to progenitor of cortical neurons |
| url | http://europepmc.org/articles/PMC4697857?pdf=render |
| work_keys_str_mv | AT irmalydiagarciacastro markersofpluripotencyinhumanamnioticepithelialcellsandtheirdifferentiationtoprogenitorofcorticalneurons AT guadalupegarcialopez markersofpluripotencyinhumanamnioticepithelialcellsandtheirdifferentiationtoprogenitorofcorticalneurons AT danielaavilagonzalez markersofpluripotencyinhumanamnioticepithelialcellsandtheirdifferentiationtoprogenitorofcorticalneurons AT hectorfloresherrera markersofpluripotencyinhumanamnioticepithelialcellsandtheirdifferentiationtoprogenitorofcorticalneurons AT anayansimolinahernandez markersofpluripotencyinhumanamnioticepithelialcellsandtheirdifferentiationtoprogenitorofcorticalneurons AT wendyportillo markersofpluripotencyinhumanamnioticepithelialcellsandtheirdifferentiationtoprogenitorofcorticalneurons AT evaramongallegos markersofpluripotencyinhumanamnioticepithelialcellsandtheirdifferentiationtoprogenitorofcorticalneurons AT nestorfabiandiaz markersofpluripotencyinhumanamnioticepithelialcellsandtheirdifferentiationtoprogenitorofcorticalneurons |