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Human multiple myeloma cells stimulate differentiation of endothelial cells to form capillary-like networks in two different co-culture systems

WANG Ya-dan HU Yu SUN Chun-yan HE Wen-juan ZHANG Xiao-ping Institution of Hematology,Affiliated Union Hospital Tongji Medical College,Huazhong University of Science and Technology,Wuhan 430022,China  
Objective To study the influence of multiple myeloma cells on differentiation of endo- thelial cells to form capillary-like networks and the role of brain derived neurotrophic factor(BDNF)in this process.Methods Human muhiple myeloma cell line RPMI8226 or fresh myeloma cells were co-cultured with human umbilical vein endothelial cells(HUVECs)in two different systems:the contact and the non-con- tact systems.The HUVECs cultured alone were used as control.The effects of soluble cytokine and adhesion molecule on angiogenesis in HUVECs co-cultured with MM cell were studied by modified Matrigel capillary- like networks formation assay and BDNF levels in co-culture system supernatant by enzyme-linked immunosor- bent assay.In the contact co-culture system,the formation of capillary-like networks and the secretion of BDNF were detected again after MM cell was preincubated and cultured with anti-CD29 and anti-CD18 mono- clonal antibodies.Results In RPMI8226 co-cuhure system,the number of capillary-like structure was in- creased in non-contact HUVECs system compared with that in monocuhure,but the increase was lower than that of contact HUVECs system(75% vs.113%).In fresh myeloma cells co-culture system,the numbers of capillary-like structure were 138% and 188% for the non-contact and the contact co-culture systems,respec- tively,above that in HUVECs cultured alone.The median BDNF concentrations in culture supernatants of monocultured,contact co-cultured with RPMI8226,non-contact co-cultured with RPMI8226,contact co-cul- tured with fresh myeloma cells and non-contact co-cultured with fresh myeloma cells were(12.4±5.1)ng/ ml,(38.5±8.2)ng/ml,(31.6±7.2)ng/ml,(37.1±8.7)ng/ml and(27.9±7.6)ng/ml,respectively. Either of the two adhesion molecule momoclonal antibodies reduced the capillary-like networks formation and the BDNF secretion in the contact co-culture system.Conclusion Human multiple myeloma cells stimulate differentiation of endothelial cells to form capillary-like networks in two different co-culture systems.BDNF takes part in this progress and is modulated by soluble cytokine and adhesion molecule expressd by both kinds of cells.
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