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Isolation of Functional Human Endothelial Cells from Small Volumes of Umbilical Cord Blood

Kang, Sa Do, Carlon, Tim A, Jantzen, Alexandra E, Lin, Fu-Hsiung, Ley, Melissa M, Allen, Jason, Stabler, Thomas V, Haley, NR, Truskey, George A and Achneck, Hardean E (2013) Isolation of Functional Human Endothelial Cells from Small Volumes of Umbilical Cord Blood. Annals of Biomedical Engineering, 41 (10). pp. 2181-2192. ISSN 0090-6964 (print) 1573-9686 (online)

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Abstract

Endothelial cells (ECs) isolated from endothelial progenitor cells in blood have great potential as a therapeutic tool to promote vasculogenesis and angiogenesis and treat cardiovascular diseases. However, current methods to isolate ECs are limited by a low yield with few colonies appearing during isolation. In order to utilize blood-derived ECs for therapeutic applications, a simple method is needed that can produce a high yield of ECs from small volumes of blood without the addition of animal-derived products. For the first time, we show that human ECs can be isolated without the prior separation of blood components through the technique of diluted whole blood incubation (DWBI) utilizing commercially available human serum. We isolated ECs from small volumes of blood (~10 mL) via DWBI and characterized them with flow cytometry, immunohistochemistry, and uptake of DiI-labeled acetylated low density lipoprotein (DiI-Ac-LDL). These ECs are functional as demonstrated by their ability to form tubular networks in Matrigel, adhere and align with flow under physiological fluid shear stress, and produce increased nitric oxide under fluid flow. An average of 7.0 ± 2.5 EC colonies that passed all functional tests described above were obtained per 10 mL of blood as compared to only 0.3 ± 0.1 colonies with the traditional method based on density centrifugation. The time until first colony appearance was 8.3 ± 1.2 days for ECs isolated with the DWBI method and 12 ± 1.4 days for ECs isolated with the traditional isolation method. A simplified method, such as DWBI, in combination with advances in isolation yield could enable the use of blood-derived ECs in clinical practice.

Item Type: Article
Uncontrolled Keywords: late-outgrowth endothelial progenitor cell, EPC, ECFC, EOC, shear stress, nitric oxide, human serum, platelet lysate, cell differentiation, cell isolation
Subjects: FOR Classification > 0903 Biomedical Engineering
FOR Classification > 1102 Cardiorespiratory Medicine and Haematology
Faculty/School/Research Centre/Department > College of Sports and Exercise Science
Depositing User: Ms Julie Gardner
Date Deposited: 23 Sep 2014 06:00
Last Modified: 13 Sep 2019 05:02
URI: http://vuir.vu.edu.au/id/eprint/25744
DOI: https://doi.org/10.1007/s10439-013-0807-5
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Citations in Scopus: 10 - View on Scopus

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