Neural fate commitment of rat full-term amniotic fluid stem cells via three-dimensional embryoid bodies and neurospheres formation

Full-term amniotic fluid stem cell (AFSC) is an underexplored reserve of broadly multipotent stem cells with potential applications in cell replacement therapy. One aspect worth exploring is the potential of AFSCs to differentiate into neural lineages. Previously, we have shown that full-term AFSC l...

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Main Authors: Azmi, Nuratiqah, Mustaffa Al Bakri, Siti Sarah, Khor, Winnie, Hamzah, Siti Nurusaadah, Ferdaos, Nurfarhana, Ling, King-Hwa, Nordin, Norshariza
Format: Article
Language:English
Published: Elsevier 2023
Online Access:http://psasir.upm.edu.my/id/eprint/108769/
http://psasir.upm.edu.my/id/eprint/108769/1/Neural%20fate%20commitment%20of%20rat%20full-term%20amniotic%20fluid.pdf
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author Azmi, Nuratiqah
Mustaffa Al Bakri, Siti Sarah
Khor, Winnie
Hamzah, Siti Nurusaadah
Ferdaos, Nurfarhana
Ling, King-Hwa
Nordin, Norshariza
author_facet Azmi, Nuratiqah
Mustaffa Al Bakri, Siti Sarah
Khor, Winnie
Hamzah, Siti Nurusaadah
Ferdaos, Nurfarhana
Ling, King-Hwa
Nordin, Norshariza
author_sort Azmi, Nuratiqah
building UPM Institutional Repository
collection Online Access
description Full-term amniotic fluid stem cell (AFSC) is an underexplored reserve of broadly multipotent stem cells with potential applications in cell replacement therapy. One aspect worth exploring is the potential of AFSCs to differentiate into neural lineages. Previously, we have shown that full-term AFSC lines established from term gestation amniotic fluid, known as R3 and R2, differentiated into neural lineage through the monolayer adherent method suggesting their neurogenic potential. The neural commitment of the cells through the formation of multicellular aggregates has never been shown before. Here, we explored the ability of R3 to commit to neural fate via the formation of three-dimensional multicellular aggregates, namely embryoid bodies (EBs) and neurospheres, exhibiting distinct characteristics resembling EBs and neurospheres as obtained from other published pluripotent and neural stem cells (NSCs), respectively. Different cell seeding densities of the cells cultured in their respective induction medium generated two distinct types of aggregates with the appropriate sizes for EBs (300–350 µm) and neurospheres (50–100 µm). The neurospheres expressed a significantly high level of Nestin than EBs. However, EBs stained positive for TUJ1, suggesting the presence of early post-mitotic neurons representing the ectodermal lineage. In contrast, the presence of the NSC population in neurosphere culture was validated with positive expression of Sox1. Notably, dissociated cells from both aggregates differentiated into MAP2-positive neural cells, highlighting the ability of both types of multicellular aggregates to commit to the neural fate. In conclusion, this study highlights the first evidence of neurosphere formation from full-term AFSCs in addition to neural fate commitment via EBs formation. Findings from this study allow researchers to select the suitable approach for neural cell generation and expansion according to research needs.
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spelling upm-1087692024-10-11T08:20:00Z http://psasir.upm.edu.my/id/eprint/108769/ Neural fate commitment of rat full-term amniotic fluid stem cells via three-dimensional embryoid bodies and neurospheres formation Azmi, Nuratiqah Mustaffa Al Bakri, Siti Sarah Khor, Winnie Hamzah, Siti Nurusaadah Ferdaos, Nurfarhana Ling, King-Hwa Nordin, Norshariza Full-term amniotic fluid stem cell (AFSC) is an underexplored reserve of broadly multipotent stem cells with potential applications in cell replacement therapy. One aspect worth exploring is the potential of AFSCs to differentiate into neural lineages. Previously, we have shown that full-term AFSC lines established from term gestation amniotic fluid, known as R3 and R2, differentiated into neural lineage through the monolayer adherent method suggesting their neurogenic potential. The neural commitment of the cells through the formation of multicellular aggregates has never been shown before. Here, we explored the ability of R3 to commit to neural fate via the formation of three-dimensional multicellular aggregates, namely embryoid bodies (EBs) and neurospheres, exhibiting distinct characteristics resembling EBs and neurospheres as obtained from other published pluripotent and neural stem cells (NSCs), respectively. Different cell seeding densities of the cells cultured in their respective induction medium generated two distinct types of aggregates with the appropriate sizes for EBs (300–350 µm) and neurospheres (50–100 µm). The neurospheres expressed a significantly high level of Nestin than EBs. However, EBs stained positive for TUJ1, suggesting the presence of early post-mitotic neurons representing the ectodermal lineage. In contrast, the presence of the NSC population in neurosphere culture was validated with positive expression of Sox1. Notably, dissociated cells from both aggregates differentiated into MAP2-positive neural cells, highlighting the ability of both types of multicellular aggregates to commit to the neural fate. In conclusion, this study highlights the first evidence of neurosphere formation from full-term AFSCs in addition to neural fate commitment via EBs formation. Findings from this study allow researchers to select the suitable approach for neural cell generation and expansion according to research needs. Elsevier 2023-06 Article PeerReviewed text en http://psasir.upm.edu.my/id/eprint/108769/1/Neural%20fate%20commitment%20of%20rat%20full-term%20amniotic%20fluid.pdf Azmi, Nuratiqah and Mustaffa Al Bakri, Siti Sarah and Khor, Winnie and Hamzah, Siti Nurusaadah and Ferdaos, Nurfarhana and Ling, King-Hwa and Nordin, Norshariza (2023) Neural fate commitment of rat full-term amniotic fluid stem cells via three-dimensional embryoid bodies and neurospheres formation. IBRO Neuroscience Reports, 14. pp. 235-243. ISSN 2667-2421 https://linkinghub.elsevier.com/retrieve/pii/S2667242123000039 10.1016/j.ibneur.2023.01.003
spellingShingle Azmi, Nuratiqah
Mustaffa Al Bakri, Siti Sarah
Khor, Winnie
Hamzah, Siti Nurusaadah
Ferdaos, Nurfarhana
Ling, King-Hwa
Nordin, Norshariza
Neural fate commitment of rat full-term amniotic fluid stem cells via three-dimensional embryoid bodies and neurospheres formation
title Neural fate commitment of rat full-term amniotic fluid stem cells via three-dimensional embryoid bodies and neurospheres formation
title_full Neural fate commitment of rat full-term amniotic fluid stem cells via three-dimensional embryoid bodies and neurospheres formation
title_fullStr Neural fate commitment of rat full-term amniotic fluid stem cells via three-dimensional embryoid bodies and neurospheres formation
title_full_unstemmed Neural fate commitment of rat full-term amniotic fluid stem cells via three-dimensional embryoid bodies and neurospheres formation
title_short Neural fate commitment of rat full-term amniotic fluid stem cells via three-dimensional embryoid bodies and neurospheres formation
title_sort neural fate commitment of rat full-term amniotic fluid stem cells via three-dimensional embryoid bodies and neurospheres formation
url http://psasir.upm.edu.my/id/eprint/108769/
http://psasir.upm.edu.my/id/eprint/108769/
http://psasir.upm.edu.my/id/eprint/108769/
http://psasir.upm.edu.my/id/eprint/108769/1/Neural%20fate%20commitment%20of%20rat%20full-term%20amniotic%20fluid.pdf