Department of Plastic and Hand Surgery, University of Freiburg Medical Center, Freiburg, Germany.
Tissue Eng Part A. 2013 Jan;19(1-2):166-74. doi: 10.1089/ten.TEA.2011.0699. Epub 2012 Sep 24.
One of the current challenges in the field of adipose tissue engineering is to promote sufficient vascularization to prevent cell death and to support adipose tissue formation. Thus, a novel strategy to enhance neovascularization of tissue-engineered adipose tissue might be the coimplantation of adipose-derived stem cells (ASCs) with endothelial progenitor cells (EPCs). However, no knowledge is given about the cellular interaction in vitro of human ASCs derived from subcutaneous fat tissue and EPCs derived from human peripheral blood. In this study, the first aim was to characterize ASCs and EPCs. Secondly, the two-dimensional Matrigel assay and the three-dimensional spheroid sprouting assay were applied for analyzing the ASC-EPC interaction in regard to formation of capillary-like structures by EPCs by ASC-conditioned medium (CM) or coculture of both cell types and compared to cocultures of ASCs and human umbilical vein endothelial cells (HUVECs). ASC-CM had no influence on the formation of capillary-like structures by EPCs. However, coculture with ASCs significantly enhanced the formation of capillary-like structures by EPCs; an effect that was not observed in cocultures of ASCs with HUVECs. Importantly, this increase in capillary-like structure formation by EPCs due to cell-cell contact was associated with significantly increased vascular endothelial growth factor (VEGF) secretion and VEGF-A mRNA expression, while inhibition of VEGF receptor tyrosine kinases completely abolished this effect. In conclusion, these data suggest that cellular communication occurs between ASCs and EPCs triggered by cell-cell contact or at least close proximity, which is partially mediated by secreted VEGF leading to the enhancement of angiogenic properties in EPCs, but not in HUVECs.
目前脂肪组织工程领域面临的挑战之一是促进足够的血管生成,以防止细胞死亡并支持脂肪组织的形成。因此,增强组织工程化脂肪组织新生血管的一种新策略可能是将脂肪来源的干细胞(ASCs)与内皮祖细胞(EPCs)共植入。然而,目前尚不清楚源自皮下脂肪组织的人 ASC 和源自人外周血的 EPC 之间在体外的细胞相互作用。在这项研究中,首先目的是对 ASC 和 EPC 进行特征描述。其次,应用二维 Matrigel 测定法和三维球体发芽测定法分析 ASC-EPC 相互作用,即 EPC 用 ASC 条件培养基(CM)或两种细胞类型的共培养形成毛细血管样结构,并与 ASC 和人脐静脉内皮细胞(HUVECs)的共培养进行比较。ASC-CM 对 EPC 形成毛细血管样结构没有影响。然而,与 ASC 的共培养显著增强了 EPC 形成毛细血管样结构的能力;而在 ASC 与 HUVEC 的共培养中则没有观察到这种作用。重要的是,由于细胞-细胞接触,EPC 形成毛细血管样结构的这种增加与血管内皮生长因子(VEGF)分泌和 VEGF-A mRNA 表达的显著增加有关,而 VEGF 受体酪氨酸激酶的抑制则完全消除了这种作用。总之,这些数据表明,ASC 和 EPC 之间发生了细胞间通讯,这种通讯是由细胞-细胞接触或至少接近触发的,部分是由分泌的 VEGF 介导的,导致 EPC 中血管生成特性增强,但在 HUVEC 中则没有。