Isolation of functional human endothelial cells from small volumes of umbilical cord blood.

Journal Article

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.

Full Text

Duke Authors

Cited Authors

  • Kang, SD; Carlon, TA; Jantzen, AE; Lin, F-H; Ley, MM; Allen, JD; Stabler, TV; Haley, NR; Truskey, GA; Achneck, HE

Published Date

  • October 2013

Published In

Volume / Issue

  • 41 / 10

Start / End Page

  • 2181 - 2192

PubMed ID

  • 23604849

Electronic International Standard Serial Number (EISSN)

  • 1573-9686

Digital Object Identifier (DOI)

  • 10.1007/s10439-013-0807-5

Language

  • eng

Conference Location

  • United States