. Author manuscript; available in PMC: 2012 Jun 1.

Published in final edited form as: Curr Diab Rep. 2011 Jun;11(3):193–202. doi: 10.1007/s11892-011-0182-2

Table 2.

Studies for ex vivo evaluation of diabetic vascular inflammation and atherosclerosis

End points examined	Design	Cell type	Findings	Limitations of method	Reference
Apoptosis and vascular injury	-Immunomagnetic isolation and staining of CD146 ⁺ vWF⁺, UEA-1⁺ CD133-cells -Syto16⁺CD45-CD31⁺CD146⁺ cells isolated by flow cytometry or by a fluorescence-activated cell sorter and electron microscopy imaging	CECs	-Opposite correlation with HbA_1c in T1DM and T2DM -Increased in diabetes -Have apoptotic cell features	-Incomplete separation from EPCs -Low circulating levels -No functional or biochemical studies -No single marker to identify -Site of endothelial damage unknown -The correlation with HbA_1c is unclear	[23-30] [31••] [32]
Apoptosis and vascular injury	-Flow cytometry analysis of platelet-poor plasma, apoptotic microparticles by Annexin V staining -TF-dependent procoagulant assay, ELISA, or prothrombinase assay -EC identity based on CD31/CD144 expression	EMPs	-Involved in cardiovascular diseases, inflammation, and coagulation -Cardiovascular risk marker in diabetes	-Incomplete standardization -Low circulating levels -No functional or biochemical studies -In diabetes, released by large panel of cells	[33-37]
Adhesion and vascular regeneration	-Cells with CD34⁺ or CD133⁺ and with KDR⁺ or CD31⁺ isolated by flow cytometry -EPC ex vivo cell culture protocol (≥ 14 days) in endothelial differentiation medium	EPCs	-Reduced or impaired in cardiovascular disease and diabetes -Form colonies, adhere to a monolayer of ECs in culture -Diabetic PBMCs can be differentiated into EPCs	-Incomplete separation from CECs/HSCs -Poor or incomplete standardization, -No functional/biochemical studies -Semiquantitative analysis -Low circulating levels -Extrapolation from in vivo environment	[38-40] [41••]
Adhesion, apoptosis, AGE/RAGE and ROS, endothelial activation/dysfunction, vascular inflammation/injury	-CD146+ ECs collected by angiocatheterization and enriched by immunomagnetic isolation -RNA amplified for quantitative PCR and microarray analysis -ECs fixed and then stained with vWF⁺ and desired markers for protein analysis	human aortic ECs/ human venous ECs	-Patients own aorta and/or venous ECs are sampled and showed ECs activation in chronic inflammation (e.g., in diabetes, cardiovascular diseases, sleep apnea, lupus and aging)	-Sampling of aorta replaced by venous for minimally invasive and repeatable sampling -Venous ECs give important, but incomplete information -RNA amplification needed for quantitative analysis	[42-44] [45] [46-49]

AGE advanced glycation end products; CECs circulating endothelial cells; ECs endothelial cells; ELISA enzyme-linked immunosorbent assay; EMPs endothelial microparticles; EPCs endothelial progenitor cells; HbA1c hemoglobin A1c; HSCs hematopoietic stem cells; PBMCs peripheral blood mononuclear cells; PCR polymerase chain reaction; RAGE receptor for advanced glycation end products; ROS reactive oxygen species; T1DM type 1 diabetes; T2DM type 2 diabetes; νWF von Willebrand factor.