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Molecular characterization of bovine amniotic fluid derived stem cells with an underlying focus on their comparative neuronal potential at different passages.
Annals of Anatomy ( IF 2.0 ) Pub Date : 2019-11-26 , DOI: 10.1016/j.aanat.2019.151452
Shah Nawaz 1 , Özlem Özden Akkaya 1 , Tayfun Dikmen 1 , Korhan Altunbaş 1 , Artay Yağci 1 , A S M Golam Kibria 2 , Metin Erdoğan 3 , Hacı Ahmet Çelik 4
Affiliation  

BACKGROUND The excellence in the field of stem cell therapy demands alternative and more convenient stem cells for potential applications. Researchers have opted for least invasive and broadly multipotent cells with minimum ethical concerns. Bovine amniotic fluid derived mesenchymal stem cells (BAF-MSCs) due to their ease of collection and owing similar gestational length to that of human could be presumed as an attractive large animal model for biomedical and biotechnology research. METHODS Bovine amniotic fluid derived stem cells were isolated from abattoir based samples and characterized for epithelial, neuronal, mesenchymal and pluripotent markers by qPCR and immunofluorescence studies at P1, P3, P5 and P7 alongside population doubling time, growth curve and multilineage differentiation studies. RESULTS The cells were explored for unique expression of Sox2, which was observed to be up regulated with increase in passage number and Nestin was found to be downregulated during further passaging of mesenchymal cells in this study. The cells also co-expressed Oct ¾ at initial passages which diminished within further passages. Evidence regarding diversity and heterogeneity in different cell population in amniotic fluid was recorded by positive expression of epithelial cell markers like pan Cytokeratin and p63 during early passages. The study suggested that cells with higher expression of Sox2 generated comparatively larger neurospheres with comparative strong expression of Sox2 and Nestin by immunofluorescence staining and qPCR analysis. Besides BAF-MSCs derived neurospheres were also shown to express pro-neuronal markers like ß-III Tubulin, GAP43 and ASCL-1. CONCLUSIONS This study explores and characterizes BAF-MSCs for their multipotent and neurogenic potentials and their use for clinical applications, though more detailed studies are needed to determine the exact pathways linked with neurogenic capacities of these cells and their morphological assessments at different gestational ages in bovines. The knowledge from the bovine model after detailed studies, proven safety and efficacy could also be used to understand substitutive strategies to investigate MSCs physiology at different trimesters and potential application of these cells for human and veterinary regenerative medicine provided the animal ethics are carefully monitored.

中文翻译:

牛羊水衍生干细胞的分子表征,其潜在的重点是它们在不同传代中的比较神经元潜能。

背景技术在干细胞治疗领域的卓越表现要求替代和更方便的干细胞用于潜在的应用。研究人员选择了具有最小伦理问题的侵入性最小的多能干细胞。牛羊水来源的间充质干细胞(BAF-MSCs)由于其易于收集和与人相似的孕期而被认为是一种有吸引力的大型动物模型,用于生物医学和生物技术研究。方法从屠宰场样本中分离牛羊水来源的干细胞,并通过qPCR和P1,P3,P5和P7免疫荧光研究,上皮细胞,神经元,间充质和多能标志物进行表征,并进行群体倍增时间,生长曲线和多系分化研究。结果研究了该细胞中Sox2的独特表达,发现该表达随传代次数的增加而上调,而Nestin在间充质细胞进一步传代期间被下调。细胞还在最初的传代中共表达Oct¾,在进一步的传代中减少。在早期传代过程中,通过上皮细胞标记物如泛细胞角蛋白和p63的阳性表达,记录了羊水中不同细胞群体的多样性和异质性的证据。研究表明,通过免疫荧光染色和qPCR分析,具有较高Sox2表达的细胞可产生较大的神经球,具有相对较强的Sox2和Nestin表达。除了BAF-MSC衍生的神经球还被证明表达前神经标记,例如ß-IIITubulin,GAP43和ASCL-1。结论这项研究探讨了BAF-MSCs的多能性和神经源性潜力,并将其用于临床应用,尽管还需要更详细的研究来确定与这些细胞的神经源性能力有关的确切途径以及在不同胎龄的牛的形态学评估。 。经过详细研究的牛模型知识,经过验证的安全性和有效性也可用于了解替代策略,以研究不同孕期MSC的生理特性,以及这些细胞在人类和兽医再生医学中的潜在应用,前提是要严格监控动物的伦理道德。结论这项研究探讨了BAF-MSCs的多能性和神经源性潜力,并将其用于临床,尽管还需要更详细的研究来确定与这些细胞的神经源性能力有关的确切途径以及它们在不同胎龄的牛的形态学评估。 。经过详细研究的牛模型知识,经过验证的安全性和有效性也可用于了解替代策略,以研究不同孕期MSC的生理特性,以及这些细胞在人类和兽医再生医学中的潜在应用,前提是要严格监控动物的伦理道德。结论这项研究探讨了BAF-MSCs的多能性和神经源性潜力,并将其用于临床应用,尽管还需要更详细的研究来确定与这些细胞的神经源性能力有关的确切途径以及在不同胎龄的牛的形态学评估。 。经过详细研究的牛模型知识,经过验证的安全性和有效性也可用于了解替代策略,以研究不同孕期MSC的生理特性,以及这些细胞在人类和兽医再生医学中的潜在应用,前提是要严格监控动物的伦理道德。尽管需要更详细的研究来确定与这些细胞的神经生成能力有关的确切途径,以及它们在不同胎龄的牛的形态学评估。经过详细研究的牛模型知识,经过验证的安全性和有效性也可用于了解替代策略,以研究不同孕期MSC的生理特性,以及这些细胞在人类和兽医再生医学中的潜在应用,前提是要严格监控动物的伦理道德。尽管需要更详细的研究来确定与这些细胞的神经生成能力有关的确切途径,以及它们在不同胎龄的牛的形态学评估。经过详细研究的牛模型知识,经过验证的安全性和有效性也可用于了解替代策略,以研究不同孕期MSC的生理特性,以及这些细胞在人类和兽医再生医学中的潜在应用,前提是要严格监控动物的伦理道德。
更新日期:2019-11-01
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