Azmi Nuratiqah, Mustaffa Al Bakri Siti Sarah, Khor Winnie, Hamzah Siti Nurusaadah, Ferdaos Nurfarhana, Ling King-Hwa, Nordin Norshariza
Department of Biomedical Science, Faculty of Medicine and Health Sciences, Universiti Putra Malaysia, 43400 Serdang, Selangor, Malaysia.
Genetics and Regenerative Medicine (ReGEN) Research Group, Faculty of Medicine and Health Sciences, Universiti Putra Malaysia, 43400 Serdang, Selangor, Malaysia.
IBRO Neurosci Rep. 2023 Jan 18;14:235-243. doi: 10.1016/j.ibneur.2023.01.003. eCollection 2023 Jun.
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 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 EBs formation. Findings from this study allow researchers to select the suitable approach for neural cell generation and expansion according to research needs.
足月羊水干细胞（AFSC）是一种尚未得到充分探索的多能干细胞储备，在细胞替代治疗中具有潜在应用价值。一个值得探索的方面是AFSCs分化为神经谱系的潜力。此前，我们已经表明，从足月妊娠羊水中建立的足月AFSC系，即R3和R2，通过单层贴壁法分化为神经谱系，表明它们具有神经发生潜力。细胞通过形成多细胞聚集体向神经方向分化此前从未被证实过。在这里，我们探索了R3形成三维多细胞聚集体（即胚状体（EBs）和神经球）并向神经命运分化的能力，这些聚集体分别表现出与其他已发表的多能干细胞和神经干细胞（NSCs）所获得的EBs和神经球相似的独特特征。在各自的诱导培养基中培养的细胞，不同的细胞接种密度产生了两种不同类型的聚集体，其大小分别适合EBs（300 - 350 µm）和神经球（50 - 100 µm）。神经球中Nestin的表达水平明显高于EBs。然而，EBs对TUJ1染色呈阳性，表明存在代表外胚层谱系的早期有丝分裂后神经元。相比之下，神经球培养中NSC群体的存在通过Sox1的阳性表达得到验证。值得注意的是，来自两种聚集体的解离细胞都分化为MAP2阳性神经细胞，突出了两种类型的多细胞聚集体向神经命运分化的能力。总之，本研究除了强调EBs形成这一神经命运分化外，还首次证明了足月AFSCs可形成神经球。本研究结果使研究人员能够根据研究需要选择合适的方法来生成和扩增神经细胞。
