Department of Obstetrics and Gynaecology, Medical University of Graz, Graz, Austria.
Institute of Pathology, Medical University of Graz, Graz, Austria.
Histochem Cell Biol. 2019 Nov;152(5):377-390. doi: 10.1007/s00418-019-01815-2. Epub 2019 Sep 21.
Function and dysfunction of endothelial cells are regulated by a multitude of factors. Endothelial cell research often requires in vitro cell culture experiments. Hence, various culture media specifically designed to promote endothelial cell growth are available. These strikingly differ in their composition: complex media contain endothelial cell growth supplement (ECGS), an extract produced of bovine brain with undefined amounts of biologically active compounds, whilst defined media contain selected growth factors in defined concentrations. We here compared the effect of seven purchasable endothelial cell culture media on colony outgrowth, proliferation, viability, in vitro angiogenesis and phenotype of mature primary human endothelial cells using feto-placental endothelial cells isolated from chorionic arteries (fpEC). The effect of media on colony outgrowth was additionally tested on umbilical cord blood-derived endothelial progenitor cells (ECFCs). Outgrowth, purity, proliferation and viability differed between media. Outgrowth of fpEC and ECFCs was best in a defined medium containing EGF, FGF2 and VEGF. By contrast, established fpEC isolations proliferated best in complex media containing ECGS, heparin and ascorbic acid. Also viability of cells was higher in complex media. In vitro angiogenesis was most intense in a defined medium containing the highest number of individual growth factors. FACS analysis of surface markers for endothelial cell subtypes revealed that endothelial phenotype of fpEC was unaffected by media composition. Our data demonstrate the fundamental effect of endothelial cell culture media on primary cell isolation success and behaviour. Whether the composition of supplements is suitable also for individual experiments needs to be tested specifically.
内皮细胞的功能和失调受多种因素调节。内皮细胞研究通常需要体外细胞培养实验。因此，有多种专门设计用于促进内皮细胞生长的培养基可供选择。这些培养基在组成上有很大的不同：复合培养基含有内皮细胞生长补充剂（ECGS），这是一种从牛脑中提取的、生物活性化合物含量不确定的提取物，而定义明确的培养基则含有选定的生长因子，浓度明确。我们在这里比较了七种可购买的内皮细胞培养基于胎盘中动脉分离的人胎盘中皮细胞（fpEC）的集落生长、增殖、活力、体外血管生成和成熟表型的影响。还在脐血衍生的内皮祖细胞（ECFCs）上测试了培养基对集落生长的影响。培养基对集落生长、纯度、增殖和活力的影响不同。含 EGF、FGF2 和 VEGF 的定义明确的培养基中 fpEC 和 ECFCs 的生长最好。相比之下，在含有 ECGS、肝素和抗坏血酸的复合培养基中，成熟的 fpEC 分离物增殖最好。在复合培养基中，细胞活力也更高。含有最多单个生长因子的定义明确的培养基中，体外血管生成最为强烈。表面标志物的 FACS 分析显示，fpEC 的内皮细胞表型不受培养基组成的影响。我们的数据表明，内皮细胞培养物对原代细胞分离成功和行为有基本影响。补充剂的组成是否也适合特定的实验，需要具体测试。
