Isolation and characterization of microvascular endothelial cells from the adult human dermis and from skin biopsies of patients with systemic sclerosis.
BACKGROUND
Systemic sclerosis (SSc) is a rheumatic autoimmune disease without known etiology and pathogenesis. Inflammatory processes with selective microvascular involvement seem to play an important role in the early stages of SSc.


METHODS
To elucidate the pathogenesis of the selective microvascular involvement in SSc, we have isolated microvascular endothelial cells from the adult human dermis (ADMEC) and for the first time from skin biopsies of patients with SSc (SSc-ADMEC) and established in cell culture. Ulex europaeus I-coated magnetic Dynabeads and the perfusion digestion technique were applied for endothelial cell isolation.


RESULTS
The cultured ADMEC and SSc-ADMEC showed the endothelial cell-specific antigenic determinants of intracellular von Willebrand factor and platelet endothelial cell adhesion molecule-1 along their cell-cell borders. These cells displayed specific uptake of acetylated low-density lipoprotein. Normal ADMEC were additionally characterized for angiotensin I-converting enzyme activity. Tumor necrosis factor-alpha activated normal ADMEC and SSc-ADMEC expressed the inflammatory adhesion molecules E-selectin, intercellular adhesion molecule-1, and vascular cell adhesion molecule-1, similarly to tumor necrosis factor-alpha-activated large-vessel endothelium represented by human umbilical cord vein endothelial cells. Normal ADMEC and human umbilical cord vein endothelial cells also expressed beta 1- and beta 4-integrin receptors in cell culture.


CONCLUSIONS
Normal ADMEC and SSc-ADMEC express endothelial cell-specific antigenic and biochemical determinants in vitro. SSc-ADMEC were for the first time established in cell culture.
Children's Hospital Department of Surgery, Boston, Massachusetts.
B M
BM
Kräling
S A
SA
Jimenez
T
T
Sorger
G G
GG
Maul
D000328:Adult; D015815:Cell Adhesion Molecules / Q000378:metabolism*; D002455:Cell Division; D002469:Cell Separation / Q000379:methods; D002478:Cells, Cultured; D004730:Endothelium, Vascular / Q000166:cytology*; D006801:Humans; D066298:In Vitro Techniques; D016023:Integrins / Q000378:metabolism*; D037102:Lectins; D008077:Lipoproteins, LDL / Q000378:metabolism; D037121:Plant Lectins; D012595:Scleroderma, Systemic / Q000473:pathology*; D012867:Skin / Q000098:blood supply / Q000166:cytology*
D015815:Cell Adhesion Molecules; D016023:Integrins; D037102:Lectins; D008077:Lipoproteins, LDL; D037121:Plant Lectins; C036850:Ulex europaeus lectins; C032629:acetyl-LDL