In Vitro Transdifferentiation Potential of Equine Mesenchymal Stem Cells into Schwann-Like Cells.

IF 2.5 3区 医学 Q3 CELL & TISSUE ENGINEERING
Lucas Vinícius de Oliveira Ferreira, Beatriz da Costa Kamura, João Pedro Marmol de Oliveira, Natielly Dias Chimenes, Marcio de Carvalho, Leandro Alves Dos Santos, Luciane Alarcão Dias-Melicio, Renée Laufer Amorim, Rogerio Martins Amorim
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引用次数: 0

Abstract

Schwann cells (SCs) are essential for the regenerative processes of peripheral nerve injuries. However, their use in cell therapy is limited. In this context, several studies have demonstrated the ability of mesenchymal stem cells (MSCs) to transdifferentiate into Schwann-like cells (SLCs) using chemical protocols or co-culture with SCs. Here, we describe for the first time the in vitro transdifferentiation potential of MSCs derived from equine adipose tissue (AT) and equine bone marrow (BM) into SLCs using a practical method. In this study, the facial nerve of a horse was collected, cut into fragments, and incubated in cell culture medium for 48 h. This medium was used to transdifferentiate the MSCs into SLCs. Equine AT-MSCs and BM-MSCs were incubated with the induction medium for 5 days. After this period, the morphology, cell viability, metabolic activity, gene expression of glial markers glial fibrillary acidic protein (GFAP), myelin basic protein (MBP), p75 and S100β, nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF), and glial cell-derived neurotrophic factor (GDNF), and the protein expression of S100 and GFAP were evaluated in undifferentiated and differentiated cells. The MSCs from the two sources incubated with the induction medium exhibited similar morphology to the SCs and maintained cell viability and metabolic activity. There was a significant increase in the gene expression of BDNF, GDNF, GFAP, MBP, p75, and S100β in equine AT-MSCs and GDNF, GFAP, MBP, p75, and S100β in equine BM-MSCs post-differentiation. Immunofluorescence analysis revealed GFAP expression in undifferentiated and differentiated cells, with a significant increase in the integrated pixel density in differentiated cells and S100 was only expressed in differentiated cells from both sources. These findings indicate that equine AT-MSCs and BM-MSCs have great transdifferentiation potential into SLCs using this method, and they represent a promising strategy for cell-based therapy for peripheral nerve regeneration in horses.

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Abstract Image

马间充质干细胞向雪旺样细胞的体外转分化潜力。
雪旺细胞(SCs)在周围神经损伤的再生过程中是必不可少的。然而,它们在细胞治疗中的应用是有限的。在这种背景下,一些研究已经证明了间充质干细胞(MSCs)通过化学方法或与SCs共培养可以转分化为雪旺样细胞(SLCs)的能力。在这里,我们首次用一种实用的方法描述了从马脂肪组织(AT)和马骨髓(BM)中提取的MSCs向SLCs的体外转分化潜力。本研究收集马面神经,切成碎片,在细胞培养基中培养48 h。使用该培养基将MSCs转分化为SLCs。马AT-MSCs和BM-MSCs在诱导培养基中孵育5 d。观察未分化和已分化细胞的形态、细胞活力、代谢活性、胶质标记物胶质纤维酸性蛋白(GFAP)、髓鞘碱性蛋白(MBP)、p75和S100β、神经生长因子(NGF)、脑源性神经营养因子(BDNF)和胶质细胞源性神经营养因子(GDNF)的基因表达,以及S100和GFAP的蛋白表达。两种来源的MSCs在诱导培养基中培养后,表现出与SCs相似的形态,并保持了细胞活力和代谢活性。分化后马AT-MSCs中BDNF、GDNF、GFAP、MBP、p75和S100β的基因表达显著增加,马BM-MSCs中GDNF、GFAP、MBP、p75和S100β的基因表达显著增加。免疫荧光分析显示,GFAP在未分化细胞和已分化细胞中均有表达,且已分化细胞的综合像元密度显著增加,S100仅在两种来源的已分化细胞中表达。这些发现表明,利用这种方法,马AT-MSCs和BM-MSCs具有很大的转分化成SLCs的潜力,它们代表了一种很有希望的基于细胞的治疗马周围神经再生的策略。
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来源期刊
Stem cells and development
Stem cells and development 医学-细胞与组织工程
CiteScore
7.80
自引率
2.50%
发文量
69
审稿时长
3 months
期刊介绍: Stem Cells and Development is globally recognized as the trusted source for critical, even controversial coverage of emerging hypotheses and novel findings. With a focus on stem cells of all tissue types and their potential therapeutic applications, the Journal provides clinical, basic, and translational scientists with cutting-edge research and findings. Stem Cells and Development coverage includes: Embryogenesis and adult counterparts of this process Physical processes linking stem cells, primary cell function, and structural development Hypotheses exploring the relationship between genotype and phenotype Development of vasculature, CNS, and other germ layer development and defects Pluripotentiality of embryonic and somatic stem cells The role of genetic and epigenetic factors in development
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