Explant combined with trypsin-digestion for culture of endothelial cells from infantile hemangiomas / 第二军医大学学报

ABSTRACT

Objective: To explore a novel method for cultivation of the endothelial cells(EC) from infantile hemangiomas (IH) in vitro and observe the biological character of cultured endothelial cells. Methods: Fresh operative specimens were obtained from infantile hemangiomas. The endothelial cells of IH were cultured by explant combined with trypsin-digestion technique. Immunohistochemical staining of EnVision method was carried out to identify the cultured cells. The purity of endothelial cell was examined by flow cytometry analysis of FITC-CD34. The biologic characters of endothelial cells were observed under inverted phase contrast microscope. Results: Endothelial cells were successfully cultured from 8 IH specimens of all the 14 explants. The morphology of cultured endothelial cells included two types:polygonal cells and fusifourm cells. The cultured cells were homogenously positive for EC markers-vWF or CD34, indicating their EC origin. Tube formation was found in endothelial cells of 1H. The proportion of CD34+ cells was 76.28% in cultured endothelial cells as detected by flow cytometry analysis. Conclusion: ECs can be successfully isolated and cultured from infantile hemangiomas by explant combined with trypsin-digestion technique; the cultured cells have some characters of endothelial cells.