A modified protocol for the isolation and culture of human umbilical vein endothelial cells

IF 2.2 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochemistry and Biophysics Reports Pub Date : 2025-09-01 DOI:10.1016/j.bbrep.2025.102221

Anmol Sandhu , Anannya Parvathi , Jennifer Jane McGhee , Salim Ismail , I-Ping Loh , Bert van der Werf , Jie Zhang , Trevor Sherwin

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Abstract

The umbilical cord is a valuable source of foetal stem cells, progenitor cells, and early-stage developmental cells, including human umbilical vein endothelial cells (HUVECs). HUVECs are widely used as a model for endothelial biology and are increasingly being investigated for their regenerative potential. Efficient isolation of these cells from the umbilical vein is a critical first step for both research and therapeutic applications. To date, most published protocols utilise Collagenase A for isolation. In this study, we present a modified HUVEC isolation protocol that employs dispase, alongside refined tissue and cell culture handling practices. We characterised the isolated cells using established HUVEC markers CD31 and CD146, and demonstrated in situ detachment of the cells from the vessel wall through immunofluorescence imaging. Our method achieved a success rate exceeding 95.6 % across all umbilical cords processed. These findings highlight the protocol's potential for broad applicability across research settings, using readily accessible reagents and equipment.

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人脐静脉内皮细胞分离和培养的改进方案

脐带是胚胎干细胞、祖细胞和早期发育细胞（包括人脐静脉内皮细胞（HUVECs））的重要来源。HUVECs被广泛用作内皮生物学模型，并因其再生潜力而受到越来越多的研究。有效地从脐静脉中分离这些细胞是研究和治疗应用的关键的第一步。迄今为止，大多数已发表的方案利用胶原酶A进行分离。在这项研究中，我们提出了一种改进的HUVEC分离方案，该方案采用了疾病，以及改进的组织和细胞培养处理方法。我们使用已建立的HUVEC标志物CD31和CD146对分离的细胞进行了表征，并通过免疫荧光成像证明了细胞与血管壁的原位分离。我们的方法在处理的所有脐带中取得了超过95.6%的成功率。这些发现突出了该方案在使用易于获得的试剂和设备的研究环境中具有广泛适用性的潜力。

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来源期刊

Biochemistry and Biophysics Reports Biochemistry, Genetics and Molecular Biology-Biophysics

CiteScore

4.60

自引率

0.00%

发文量

191

审稿时长

59 days

期刊介绍： Open access, online only, peer-reviewed international journal in the Life Sciences, established in 2014 Biochemistry and Biophysics Reports (BB Reports) publishes original research in all aspects of Biochemistry, Biophysics and related areas like Molecular and Cell Biology. BB Reports welcomes solid though more preliminary, descriptive and small scale results if they have the potential to stimulate and/or contribute to future research, leading to new insights or hypothesis. Primary criteria for acceptance is that the work is original, scientifically and technically sound and provides valuable knowledge to life sciences research. We strongly believe all results deserve to be published and documented for the advancement of science. BB Reports specifically appreciates receiving reports on: Negative results, Replication studies, Reanalysis of previous datasets.