hmvec cells  (Cell Applications Inc)

Journal: The British journal of dermatology

Article Title: Coexistence of Eph receptor B 1 and ephrin B 2 in port-wine stain endothelial progenitor cells contributes to clinicopathological vasculature dilatation

doi: 10.1111/bjd.15716

Figure Lengend Snippet: Port-wine stain (PWS) endothelial cells (ECs) showed dual arterial and venous identities of co-expression of Eph receptor B1 (EphB1) and ephrin B2 (EfnB2). (a, b) PWS ECs were EphB1+ and EfnB2+. (c, d) A normal dermal venule showed expression of EphB1 but not EfnB2. (e) PWS ECs were EphB4−. (f) Relative mRNA levels of EphB1 and EfnB2 in selected normal human dermal microvascular endothelial cell (hDMVEC) subtypes. (g–i) Forced co-expression of EphB1 and EfnB2 in normal hDMVECs showed a significant increase in (g) branch thickness, (h) branch point area and (i) tube circumference of the capillary tubes formed in vitro compared with controls. (j) PWS blood vessel-like phenotypes were observed in EphB1+/EfnB2+ but not in wild-type and EphB1+/EGFP+ control hDMVECs in an in vitro capillary tube formation assay at 12 h after cell plating. Positive stain is diaminobenzidine (DAB; brown). Scale bar = 100 μm. *P < 0·05, ** P < 0·01 vs. control. The arrows indicate blood vessels. (k) Left panel, detection of expression of EphB1, EphB4, β-actin and EfnB2 by Western blot (W.B.) in various hDMVEC subpopulations. hDMVEC, heterogeneous population prior to EfnB2–Fc selection; EphB1− hDMVEC, the remaining hDMVEC subpopulation after EfnB2–Fc selection; enhanced green fluorescent protein (EGFP) or EfnB2–His, overexpression of EGFP or EfnB2 in the EfnB2–Fc selected hDMVEC subpopulation. Right panel, an anti-EphB1 antibody was used to immunoprecipitate EphB1 from cell lysate and EfnB2 was detected from the immunoprecipitated protein complex using an anti-EfnB2 or anti-His antibody.

Article Snippet: Human dermal microvascular endothelial cells (hDMVECs) were purchased from Cell Applications (San Diego, CA, U.S.A.) and were cultured in EC Basal Medium with growth supplement (Cell Applications).

Techniques: Staining, Expressing, In Vitro, Control, Capillary Tube Formation Assay, Western Blot, Selection, Over Expression, Immunoprecipitation

Journal: The British journal of dermatology

Article Title: Coexistence of Eph receptor B 1 and ephrin B 2 in port-wine stain endothelial progenitor cells contributes to clinicopathological vasculature dilatation

doi: 10.1111/bjd.15716

Figure Lengend Snippet: Port-wine stain (PWS) endothelial cells (ECs) showed dual arterial and venous identities of co-expression of Eph receptor B1 (EphB1) and ephrin B2 (EfnB2). (a, b) PWS ECs were EphB1+ and EfnB2+. (c, d) A normal dermal venule showed expression of EphB1 but not EfnB2. (e) PWS ECs were EphB4−. (f) Relative mRNA levels of EphB1 and EfnB2 in selected normal human dermal microvascular endothelial cell (hDMVEC) subtypes. (g–i) Forced co-expression of EphB1 and EfnB2 in normal hDMVECs showed a significant increase in (g) branch thickness, (h) branch point area and (i) tube circumference of the capillary tubes formed in vitro compared with controls. (j) PWS blood vessel-like phenotypes were observed in EphB1+/EfnB2+ but not in wild-type and EphB1+/EGFP+ control hDMVECs in an in vitro capillary tube formation assay at 12 h after cell plating. Positive stain is diaminobenzidine (DAB; brown). Scale bar = 100 μm. *P < 0·05, ** P < 0·01 vs. control. The arrows indicate blood vessels. (k) Left panel, detection of expression of EphB1, EphB4, β-actin and EfnB2 by Western blot (W.B.) in various hDMVEC subpopulations. hDMVEC, heterogeneous population prior to EfnB2–Fc selection; EphB1− hDMVEC, the remaining hDMVEC subpopulation after EfnB2–Fc selection; enhanced green fluorescent protein (EGFP) or EfnB2–His, overexpression of EGFP or EfnB2 in the EfnB2–Fc selected hDMVEC subpopulation. Right panel, an anti-EphB1 antibody was used to immunoprecipitate EphB1 from cell lysate and EfnB2 was detected from the immunoprecipitated protein complex using an anti-EfnB2 or anti-His antibody.

Article Snippet: Human dermal microvascular endothelial cell culture Human dermal microvascular endothelial cells (hDMVECs) were purchased from Cell Applications (San Diego, CA, U.S.A.) and were cultured in EC Basal Medium with growth supplement (Cell Applications).

Techniques: Staining, Expressing, In Vitro, Control, Capillary Tube Formation Assay, Western Blot, Selection, Over Expression, Immunoprecipitation