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Isolation, culture and identification of human venous malformation endothelial cells
Jun Jia1, Yifang Zhao, Wenfeng Zhang
1Department of Oral and Maxillofacial Surgery, School of Stomatology, Wuhan University, Wuhan 430079, China.
Objective:
To establish a method of culturing endothelial cells (EC) from human vascular malformation.
Methods:
Venous malformation specimens obtained from the patient undergoing oral surgery were plated into glass, plastic and gelatin-coated dishes. Pure cultures of human vascular malformation endothelial cells (VMEC) were isolated by the ways of discarding the tissues at early stage of primary culture, scraping and trypsinizing. Morphological characteristics were studied under phase-contrast microscope and electron microscope (EM), and determined by immunohistochemistry.
Results:
The cells were pure and could be maintained in culture up to 4 approximately 5 passages, or 40 approximately 50 days. VMEC formed contact-inhibited "cobblestone" monolayer on glass and plastic, and capillary-like "tubes" on gelatin. EM revealed that there were Weibel-Palade bodies in the culture cells. The cells showed positive staining for CD(34), vWF, and negative for alpha-SMA.
Conclusions:
The culture technique for growing VMEC has been established. And these cells can provide a useful tool for studying biological characteristics of human vascular malformation in vitro.