Distler J H W, Allanore Y, Avouac J, Giacomelli R, Guiducci S, Moritz F, Akhmetshina A, Walker U A, Gabrielli A, Müller-Ladner U, Tyndall A, Matucci-Cerinic M, Distler O
Department for Internal Medicine 3 and Institute for Clinical Immunology, University of Erlangen-Nuremberg, Erlangen, Germany.
Ann Rheum Dis. 2009 Feb;68(2):163-8. doi: 10.1136/ard.2008.091918. Epub 2008 Jul 23.
Systemic sclerosis (SSc) is characterised by a progressive microangiopathy that contributes significantly to the morbidity of patients with SSc. Besides insufficient angiogenesis, defective vasculogenesis with altered numbers of endothelial precursor cells (EPCs) might also contribute to the vascular pathogenesis of SSc. However, different protocols for isolation, enrichment, culture and quantification of EPCs are currently used, which complicate comparison and interpretation of the results from different studies. The aim of the European League Against Rheumatism Scleroderma Trials and Research (EUSTAR) group expert panel was to provide recommendations for standardisation of future research on EPCs. Consensus statements and recommendations were developed in a face to face meeting by an expert panel of the basic science working group of EUSTAR. The findings were: cardiovascular risk factors and medications such as statins should be described in detail. A detailed description of methods considering isolation, culture, enrichment and detection of EPCs should be given. For in vitro culture of EPCs, no protocol has been shown to be superior to another, but coating with laminin and type IV collagen would resemble most closely the situation in vivo. The endothelial phenotype should be confirmed in all in vitro cultures at the end of the culture period. We recommend using CD133, vascular endothelial growth factor type 2 receptor (VEGFR2) and CD34 in combination with a viability marker for quantification of EPCs in the blood. Finally, exact standard operating procedures for fluorescence-activated cell sorting (FACS) analysis are given that should be strictly followed. In summary, the EUSTAR recommendations will help to unify EPC research and allow better comparison between the results of different studies.
系统性硬化症（SSc）的特征是进行性微血管病变，这对SSc患者的发病率有重大影响。除了血管生成不足外，内皮祖细胞（EPCs）数量改变导致的血管生成缺陷也可能在SSc的血管发病机制中起作用。然而，目前用于分离、富集、培养和定量EPCs的方案各不相同，这使得不同研究结果的比较和解释变得复杂。欧洲抗风湿病联盟硬皮病试验与研究（EUSTAR）小组专家小组的目的是为未来EPCs研究的标准化提供建议。EUSTAR基础科学工作组的专家小组在一次面对面会议上制定了共识声明和建议。研究结果如下：应详细描述心血管危险因素和他汀类药物等药物。应详细描述考虑EPCs分离、培养、富集和检测的方法。对于EPCs的体外培养，尚无一种方案被证明优于另一种，但用层粘连蛋白和IV型胶原包被最接近体内情况。在培养期结束时，应在所有体外培养物中确认内皮细胞表型。我们建议联合使用CD133、血管内皮生长因子2型受体（VEGFR2）和CD34以及一种活力标记物来定量血液中的EPCs。最后，给出了荧光激活细胞分选（FACS）分析应严格遵循的确切标准操作程序。总之，EUSTAR的建议将有助于统一EPC研究，并使不同研究结果之间具有更好的可比性。
