Isolation of Umbilical Cord-Derived Mesenchymal Stem Cells with High Yields and Low Damage

间充质干细胞 脐带 干细胞 分离(微生物学) 帘布衬里 生物 细胞生物学 成体干细胞 免疫学 生物信息学 内皮干细胞 体外 遗传学
作者
Mingyuan Xu,Jiasen Xu,Danling Cheng,Xiaoni Chen,Shan Lin,Xinlei Si,Fangfang Guo,Di Wu,Fenfang Wu
出处
期刊:Journal of Visualized Experiments [MyJoVE Corporation]
卷期号: (209)
标识
DOI:10.3791/66835
摘要

Mesenchymal stem cells (MSCs) are a population of multipotent cells with remarkable regenerative and immunomodulatory properties. Wharton's jelly (WJ) from the umbilical cord (UC) has gained increasing interest in the biomedical field as an outstanding source of MSCs. However, challenges such as limited supply and lack of standardization in existing methods have arisen. This article presents a novel method for enhancing MSC yield by dissecting intact WJ from the umbilical cord. The method employs blunt dissection to remove the epithelial layer, maintaining the integrity of the entire WJ and resulting in an increased quantity and viability of harvested MSCs. This approach significantly reduces WJ waste compared to conventional sharp dissection methods. To ensure the purity of WJ-MSCs and minimize external cellular influence, a procedure utilizing internal tension to peel off the endothelium after flipping the UC was conducted. Additionally, the Petri dish was inverted for a short time during explant culture to improve attachment and cell outgrowth. Comparative analysis demonstrated the superiority of the proposed method, showing a higher yield of WJ and WJ-MSCs with better viability than traditional methods. The similar morphology and expression pattern of cell surface markers in both methods confirm their characterization and purity for various applications. This method provides a high-yield and high-viability approach for WJ-MSC isolation, demonstrating great potential for the clinical application of MSCs.

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