Mustaffa Al Bakri, Siti Sarah (2021) Generation and characterization of amniotic fluid stem cell (AFSC) Line -derived neural stem cell ( NSC). [Project Paper] (Submitted)
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Abstract
Neural stem cell (NSC) serves as a high prospective cell for neuro-transplantation as one of the treatments for neurodegenerative diseases (NDs). The inaccessibility of the brain sourced NSCs limits its application and leads to the idea of generating NSCs from non-brain sources. Amniotic fluid stem cells (AFSCs) are neurogenic and a potential non-brain source of NSCs. Due to its accessibility and therapeutic potential, the isolation of AFSCs from full-term gestation has garnered a lot of attention, including as a potential non-brain source for NSCs. Objective: Our study aims to evaluate the proof of concept on the potential of full-term rat amniotic fluid stem cell line (R3) to generate NSCs and the ability of the R3-derived NSCs to form neurospheres and differentiate into neurons. Methodology: R3 was cultured in a medium containing GMEM supplemented with 10% foetal bovine serum (FBS), other essential supplements and 20 ng/ml leukemia inhibitory factor (LIF) prior to transdifferentiation into NSCs through monolayer differentiation (MD) assay in NSC culture medium for two days. Optimization on the NSC induction media was performed at this stage using two different NSC induction media (with serum and without serum). The generation of NSCs was then confirmed by immunocytochemistry (ICC) analysis on the expression of NSC protein markers (Nestin, Sox2). R3-derived NSCs were further tested on its ability to form multicellular aggregates, neurospheres, by plating NSCs in low attachment plates in NSC medium for 3 days. The diameter and the ability of the neurospheres to form neurons were examined using the Cell Sens Standard computer software and immunocytochemistry (ICC) for neuronal markers expression (Tuj1, MAP2), respectively. Results: NSC induction medium with serum provides optimum condition to promote transdifferentiation of R3. R3 has successfully transdifferentiated into NSCs and expressed Nestin, one of the NSC markers. Unfortunately, due to technical error, Sox2 expression could not be detected. The R3-derived NSCs successfully formed neurospheres and further differentiated into neurons expressing the neuronal markers -Tuj-1 and MAP2 by ICC analysis. Discussion: Addition of serum in the NSC induction medium provides crucial factors to promote the regulation and maintenance of neural lineage stem cells as marked by the expression of Nestin marker. Generation of neurosphere with appropriate size is crucial for adequate supply of nutrient and metabolite for viability and differentiation into neural cells. The expression of post-mitotic (Tuj-1) and mature (MAP2)- neuron markers reveal the ability of R3-derived NSCs to form neurons. Conclusion: Findings of this study signify the ability of stem cells from amniotic fluid at full gestation as a potential non-brain source of NSCs, which could prospectively be useful for treatment of NDs.
| Item Type: | Project Paper |
|---|---|
| Faculty: | Faculty of Medicine and Health Science |
| Depositing User: | Ms. Nor Safa'aton Saidin |
| Date Deposited: | 22 Aug 2023 07:49 |
| Last Modified: | 22 Aug 2023 07:49 |
| URI: | http://psaspb.upm.edu.my/id/eprint/1139 |
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