Objective: To investigate the effects of beta-blocker, propranolol and timolol on the proliferation and apoptosis of hemangioma stem cells (HemSCs). Methods: Different concentrations(3, 30, 90, 150 μmol/L)of propranolol and timolol were added in HemSCs culture respectively for 24, 48 and 72 h, and cell proliferation and apoptosis were detected by flow cytometry. Experimental data were analyzed using SPSS 19.0 software. Statistically significance was determined using Student′s t test. Values of P<0.05 were considered statistically significant. Results: Proliferation assay indicated that different concentrations of propranolol and timolol could inhibit HemSCs proliferation, and inhibition degree increased with increasing concentration. However, the effects of propranolol was slightly stronger than that of timolol at high concentrations (30, 90 and 150 μmol/L), but the opposite was observed at low concentration(3 μmol/L)(P<0.05). The results of apoptosis experiment showed that timolol had stronger effect than propranolol in promoting apoptosis at low concentrations (3 and 30 μmol/L), which was 13.7% and 1.1% higher, respectively. Conclusions Inhibiting HemSCs proliferation and promoting apoptosis may be one of the mechanisms of propranolol and timolol in treating infantile hemangioma.The specific effects of both varies with the concentration and time.

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.

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.