Kubiena H, Mädel C, Roka J, Burjak S, Frey E, Frey M
Klinische Abteilung für Plastische und Rekonstruktive Chirurgie, Universitätsklinik für Chirurgie, Medizinische Universität Wien, Osterreich.
Handchir Mikrochir Plast Chir. 2009 Apr;41(2):107-11. doi: 10.1055/s-2008-1039014. Epub 2009 Mar 3.
Haemangioma is the most common tumour of infancy. Its biological behaviour is one of a benign nature and characterised by a typical biphasic growth pattern. After an initial proliferation during the child's first year of life an involution takes place and usually lasts until the first decade. The expression of cellular and extracellular markers as well as cytokines follows its biphasic growth. In-vitro models reported so far have provided new insights in the tumour's biology and have identified possible targets for pharmacological agents.
Our study should enhance the functionality of current in-vitro models in establishing a suspension model of isolated proliferating haemangioma vascular endothelial cells (HVECs).
Tissue samples isolated from four haemangiomas (H1-H4) during the proliferation phase were cultivated in four different ways: H1 was cultivated in an MCDB-131 medium whereas for the H2 samples EGM2-MV medium was used. In the H3 series dispase enabled an immediate isolation of pure HVECs from the tissue sample which were cultivated under EMG2-MV medium. In the similarly cultivated H4 series an isolation of cultivated HVECs was enabled by the use of magnetic bead separation which finally led to a suspension model of isolated HVECs.
In all four series HVECs were cultivated successfully. Depending on the selected medium different growth rates were observed. The use of dispase and the magnetic bead separation technique resulted in an isolation of cultivated HVECs which was documented in haematoxylin/eosin staining as well as under CD-31 immunohistochemistry.
Research on tumour-specific processes as well as possible pharmacological targets necessitates a stable in-vitro model of proliferating HVECs. For our purposes, the described suspension model of HVECs fulfills the requirements of further research on antiproliferation and anti-angiogenesis.
血管瘤是婴儿期最常见的肿瘤。其生物学行为具有良性性质,特征为典型的双相生长模式。在儿童出生后的第一年经历初始增殖阶段后,会发生退化,通常持续到第一个十年。细胞和细胞外标志物以及细胞因子的表达遵循其双相生长。迄今为止报道的体外模型为肿瘤生物学提供了新的见解,并确定了药物制剂的可能靶点。
我们的研究应增强当前体外模型在建立分离的增殖性血管瘤血管内皮细胞(HVECs)悬浮模型方面的功能。
从增殖期的四个血管瘤(H1 - H4)中分离出的组织样本以四种不同方式培养:H1在MCDB - 131培养基中培养,而H2样本使用EGM2 - MV培养基。在H3系列中,胰蛋白酶能够从组织样本中立即分离出纯的HVECs,并在EMG2 - MV培养基下培养。在类似培养的H4系列中,通过使用磁珠分离实现了培养的HVECs的分离,最终形成了分离的HVECs悬浮模型。
在所有四个系列中,HVECs均成功培养。根据所选培养基观察到不同的生长速率。胰蛋白酶和磁珠分离技术的使用导致了培养的HVECs的分离,这在苏木精/伊红染色以及CD - 31免疫组织化学中得到了证实。
对肿瘤特异性过程以及可能的药理学靶点的研究需要一个稳定的增殖性HVECs体外模型。就我们的目的而言,所描述的HVECs悬浮模型满足了进一步抗增殖和抗血管生成研究的要求。