Isolation, culture and characterization of infantile hemangioma stem cell.

OBJECTIVE

To establish a reliable method of isolation and culture of infantile hemangioma stem cells (HemSCs).

METHODS

Proliferating infantile hemangioma specimens were digested with collagenase to form a single cell suspension. The HemSCs were isolated with anti-CD133 MicroBeads, and were incubated in fibronectin coated 96-well plates with EBM-2 (10％ FBS). HemSCs were identified by morphological characteristics, flow cytometry, cell tubule formation assay, osteoinductive and adipogenic differentiation assay, and subcutaneous tumor formation assay.

RESULTS

This method enables the rapid isolation of HemSCs which demonstrated typical mesenchymal stem cell morphology in culture.CD133 (+) HemSCs expressed CD29 (99.5％),CD44 (97.9％),CD90 (87.6％) and CD105 (98.5％),but barely expressed CD31 (0.2％),CD34 (0.1％),CD45 (0.1％) and CD144 (0.1％).These cells could differentiate into osteoblasts and adipocytes,and could form vascular wall like structure in vitro. When implanted into subcutaneous of the nude mice, the cells can develop into hemangioma like lesion histologically.

CONCLUSIONS

This technique can effectively isolate HemSCs from the proliferative hemangioma. These cells could be further used to reveal the charaeteristics of HemSCs, as well as for further study of widespread application.