The Potentiality of Human Umbilical Cord Isolated Mesenchymal Stem/Stromal Cells for Cardiomyocyte Generation.

IF 2.6 Q4 CELL BIOLOGY Stem Cells and Cloning-Advances and Applications Pub Date : 2020-11-09 eCollection Date: 2020-01-01 DOI:10.2147/SCCAA.S253108
Amoura Abou-ElNaga, Farha El-Chennawi, Samar Ibrahim Kamel, Ghada Mutawa
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引用次数: 5

Abstract

Background: The new therapeutic strategy of managing cardiac diseases is based on cell therapy; it highly suggests the use of multipotent mesenchymal stem/stromal cells (MSCs). MSCs widely used in researches are known to be isolated from bone marrow. However, this research seeks to use a human umbilical cord (HUC) as an alternative source of MSCs. Since HUC Wharton's jelly (WJ)-isolated MSCs originate as fetal tissue they are highly preferable for their potential advantages over other adult tissues.

Methods: The researchers used enzymatic digestion to establish a primary HUC-WJ-isolated MSC line. Then, flow cytometry was used to characterize MSCs and hematopoietic stem cells (HSCs) markers' expression. In addition, the cardiac differentiation capacity of HUC-WJ-isolated MSCs in vitro was investigated by two protocols. Protocol-1 necessitates the dependence on merely 5-azacytidine (5-Aza), whereas in protocol-2, 5-Aza was supported by basic fibroblast growth factor (BFGF). The comparative study between the two protocols was applied by inspecting the ultrastructure of differentiated cells, measuring RT-PCR mRNA cardiac markers and the quantitative detection of cardiac proteins.

Results: HUC-WJ isolated MSCs were expressed by CD90+ve, CD105+ve, CD106+ve, CD45-ve, and CD146-ve. Remarkable TNNT1, NKX2.5, and Desmin mRNA expression and higher quantitative LDH and cTnI were detected by applying protocol-2. This same protocol-2 induced cardiac morphological features that were revealed by identifying cardiomyocyte-like cells and typical sarcomeres.

Conclusion: HUC-WJ is proved to be an ethical and effective source of MSCs induced cardiac differentiation, whereas BFGF supports 5-Aza in MSCs-cardiomyocytes differentiation.

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人脐带分离间充质干细胞/基质细胞用于心肌细胞生成的潜力。
背景:心脏疾病的治疗新策略是基于细胞治疗;它强烈建议使用多能间充质干细胞(MSCs)。研究中广泛使用的间充质干细胞是从骨髓中分离出来的。然而,这项研究试图使用人类脐带(HUC)作为骨髓间充质干细胞的替代来源。由于HUC沃顿果冻(WJ)分离的间充质干细胞起源于胎儿组织,因此它们比其他成人组织具有更大的潜在优势。方法:采用酶切法建立huc - wj分离的MSC初代细胞系。然后用流式细胞术检测MSCs和造血干细胞(hsc)标志物的表达。此外,通过两种方案研究huc - wj分离的MSCs体外心脏分化能力。方案1要求仅依赖5-氮杂胞苷(5-Aza),而方案2要求碱性成纤维细胞生长因子(BFGF)支持5-Aza。通过观察分化细胞超微结构、检测心脏标志物RT-PCR mRNA和定量检测心脏蛋白,对两种方案进行比较研究。结果:HUC-WJ分离的MSCs分别表达CD90+ve、CD105+ve、CD106+ve、CD45-ve和CD146-ve。应用方案2检测到TNNT1、NKX2.5和Desmin mRNA的显著表达,LDH和cTnI的定量升高。通过鉴定心肌细胞样细胞和典型的肌瘤,同样的方案2诱导了心脏形态学特征。结论:HUC-WJ是MSCs诱导心肌细胞分化的有效来源,而BFGF支持5-Aza促进MSCs-心肌细胞分化。
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CiteScore
6.50
自引率
0.00%
发文量
10
审稿时长
16 weeks
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