Endothelial Progenitor Cells Derived from Human Umbilical Cord Blood Culture in Vitro with Differential Attachment Method / 中华高血压杂志

Objective To establish a practical,stable and high purity endothelial progenitor cells culture meth- od in vitro.Methods Human umbilical cord blood mononuclear cells were isolated by Ficoll density-gradient cen- trifugation,then plated on dishes coated with human fibronectin.After 48 hours,the nonaderent cells were collect- ed and replated onto fibronectin-coated dishes.After 7 days of culture,the cells were identified with the techniques of immunohistochemistry,immunofluorescence and flow cytometer.Results The cultured cells were small and spindle or polygonal in shape.Large numbers of typical endothelial progenitor cell colony-forming units were found,vWF and Flk-1 proteins expression were identified in more than 95% of the attached cells with 98% of them showing positive Dil-ac-LDL and FITC-UEA-1.According to the results from fluorescence-activated cell sorting (FACS),7.0%?1.8% of cells were recognized as CD_(133)~+.Conclusion Differential attachment technique is a practical and stable method for obtaining highly purified endothelial progenitor cells.

Number of circulating endothelial progenitor cells inversely correlate with the severity of coronary artery disease in patients with stable coronary artery disease / 中华心血管病杂志

<p><b>OBJECTIVE</b>To investigate whether the number of circulating endothelial progenitor cells (EPCs) correlate with the severity of coronary stenosis in patients with stable coronary artery disease (CAD).</p><p><b>METHODS</b>80 consecutive patients who underwent coronary angiography (exclusion of acute coronary syndrome and myocardial infarction) were enrolled. Physical examination and blood tests were performed to assess the disease severity and cardiovascular risk factors. Circulating EPCs as measured by the number of CD133/KDR double positive cells were detected by FACS.</p><p><b>RESULTS</b>The number of EPCs inversely correlated with age, creatinine clearance (Ccr) and left ventricular mass index (LVMI) (P = 0.004, 0.015, 0.014 respectively). Patients with hypertension showed significant reduction in number of EPCs compared to those without hypertension (P = 0.004). Moreover, the number of EPCs in patients with coronary artery disease was significantly lower than that of those with normal coronary artery (P < 0.01). EPCs also inversely correlated with angiographic Gensini score (n = 49, r = -0.305, P = 0.039).</p><p><b>CONCLUSIONS</b>In patients with stable CAD, the numbers of circulating EPCs correlate with the severity of CAD as well as cardiovascular risk factors.</p>

Osteopontin enhances migratory ability of cultured aortic adventitial fibroblasts from spontaneously hypertensive rats / 生理学报

Migration of adventitial fibroblasts (AF) is involved in the neointimal formation which is one of the common pathological processes in several vascular diseases. The observation of whether the migratory response of AF from hypertensive animal is different from that of controls may provide an explanation of vascular remodeling in hypertension. We examined whether there is any difference between the migratory activity of AF derived from spontaneously hypertensive rat (SHR) and that from their normotensive counterpart Wistar-Kyoto rats (WKY). In addition, the role of osteopontin (OPN) in cell migration was also examined. Primary cultures of aortic adventitial fibroblasts were derived from SHR and age-matched WKY. Migration of fibroblasts was determined with the Transwell method. The mRNA expression level of OPN was measured by a real-time quantitative PCR. When compared with WKY-derived cells, migration of adventitial fibroblasts from SHR exhibited an increased response when stimulated by 10% serum (cell number per field 35.20+/-5.26 vs 22.2+/-3.27, p<0.05). Chemotaxis induced by 10 ng/ml bFGF showed a similar difference (cell number per field 30.23+/-4.54 vs 19.20+/-4.47, p<0.05). We also found that SHR-derived fibroblasts expressed a higher level of OPN mRNA than the cells from WKY (1863.23+/-43.91 vs 326.24+/-68.29, p<0.01). To verify if OPN is associated with the enhanced migratory ability in AF from SHR, we designed the antisense oligonucleotide of OPN. The results showed that the antisense OPN oligonucleotide significantly inhibited AF migration (cell number per field 38.60+/-5.98 vs 26.61+/-3.84, p<0.05), while sense and mismatch OPN oligonucleotide had no effect on cell migration. Therefore, the migration of adventitial fibroblasts appeared to be enhanced in cultures derived from SHR. OPN might be involved in the difference observed.