Hofbauer Pablo, Riedl Sabrina, Witzeneder Karin, Hildner Florian, Wolbank Susanne, Groeger Marion, Gabriel Christian, Redl Heinz, Holnthoner Wolfgang
Ludwig Boltzmann Institute for Experimental and Clinical Traumatology, AUVA Research Center, Austrian Cluster for Tissue Regeneration, Vienna, Austria.
Red Cross Blood Transfusion Service of Upper Austria, Austrian Cluster for Tissue Regeneration, Linz, Austria.
Cytotherapy. 2014 Sep;16(9):1238-44. doi: 10.1016/j.jcyt.2014.04.009. Epub 2014 Jun 11.
As angiogenic and lymphangiogenic key players, endothelial cells (ECs) are promising candidates for vascular regenerative therapies. To culture ECs in vitro, fetal calf serum (FCS) is most often used. However, some critical aspects of FCS usage, such as possible internalization of xenogeneic proteins and prions, must be considered. Therefore, the aim of this project was to determine if human platelet lysate (hPL) is a suitable alternative to FCS as medium supplement for the culture of blood vascular and lymphatic endothelial cells.
The usability of hPL was tested by analysis of endothelial surface marker expression, metabolic activity and vasculogenic potential of outgrowth ECs (OECs), human umbilical vein ECs (HUVECs), and lymphatic ECs (LECs).
Expression of EC markers CD31, VEGFR2, VE-cadherin and CD146 did not differ significantly between the EC types cultured in FCS or hPL. In addition, OECs, HUVECs and LECs formed tube-like structures on Matrigel when cultured in hPL and FCS. With the use of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazoliumbromid assays, we found that the metabolic activity of OECs and LECs was slightly decreased when hPL was used. However, HUVECs and LECs did not show a significant decrease in metabolic activity, and HUVECs showed a slightly higher activity at low seeding densities.
The use of hPL on different EC types did not reveal any substantial negative effects on EC behavior. Thus, hPL appears to be a favorable candidate to replace FCS as a medium supplement in the culture of ECs.
作为血管生成和淋巴管生成的关键参与者，内皮细胞（ECs）是血管再生治疗的理想候选细胞。在体外培养内皮细胞时，最常使用胎牛血清（FCS）。然而，使用FCS时的一些关键问题，如异种蛋白和朊病毒的可能内化，必须加以考虑。因此，本项目的目的是确定人血小板裂解物（hPL）是否是FCS的合适替代品，作为血管和淋巴管内皮细胞培养的培养基补充剂。
通过分析增殖型内皮细胞（OECs）、人脐静脉内皮细胞（HUVECs）和淋巴管内皮细胞（LECs）的内皮表面标志物表达、代谢活性和血管生成潜力，测试hPL的可用性。
在FCS或hPL中培养的内皮细胞类型之间，EC标志物CD31、VEGFR2、VE-钙黏蛋白和CD146的表达没有显著差异。此外，当在hPL和FCS中培养时，OECs、HUVECs和LECs在基质胶上形成管状结构。使用3-（4,5-二甲基噻唑-2-基）-2,5-二苯基四氮唑溴盐试验，我们发现使用hPL时，OECs和LECs的代谢活性略有下降。然而，HUVECs和LECs的代谢活性没有显著下降，并且在低接种密度下，HUVECs的活性略高。
在不同的内皮细胞类型上使用hPL对内皮细胞行为没有显示出任何实质性的负面影响。因此，hPL似乎是替代FCS作为内皮细胞培养中培养基补充剂的有利候选物。
