Product Name 3D glomerular microvascular AngiogenesisCatalog Number EP005Product Format 6 ,12, and 24 wellStorage 37° C
GENERAL INFORMATIONOur 3D Human Glomerular Microvascular Angiogenesis proprietary system in which GFPtagged human endothelial cells from variable vascular beds are co-cultured with RFPtagged human cells in a specially designed medium. The endothelial cells initially formsmall islands within the culture matrix. They subsequently begin to proliferate and thenenter a migratory phase during which they move through the matrix to form threadlikestructures with lumens. These gradually join up by (1-2 weeks) to form a network ofanastomosing tubules which closely resembles the capillary bed found in vivo.The 3D Human Glomerular Microvascular Angiogenesis contains all of the materialsnecessary to perform multiple angiogenesis assays in 6, 12, or 24 well formats. The 3Dmodel is designed that the testing materials, i.e. compounds, conditioned media, or tissueexplants, can be added into the system at any time, ranging from the onset ofvasculogenesis to advanced angiogenesis. The resulting effect on tubule formation (tubularlength, number of branches et al) can be monitored throughout the whole process underinverted fluorescence microscope.Reagents and Materials Provided:1. 1 x vial of mixture of GFP-tagged Human Glomerular Microvascular ECs and RFPtagged human supporting cells (-80°C/liquid N2)2. 1 x 24-well plate coated with AlphaBiocoat solution (Room temperature, for 2months)3. 1 x 500ml of Endo-Growth Medium (4°C)
Day 11. Pre-warm Endo-Growth Medium to 37ºC in a water bath2. Accurately pipette 24ml Endo-Growth Medium into a 50ml Falcon tube;3. Rapidly thaw the vial of cryopreserved cells in a 37ºC water bath;4. Transfer all cells gently into 24ml pre warmed Endo-growth medium;5. Mix well the cells gently using a serological pipette;6. Add 1.0ml of cell suspension to each well of the pre-coated 24-well plate.7. Make sure the cells are evenly dispersed in the wells.8. Place the plate in an incubator (37ºC, 5% CO2 and humidified).Day 29. Take the plate from the incubator and examine cells under inverted fluorescencemicroscopy10.(GFP positive Human Glomerular Microvascular ECs should sparsely and evenlydistributed among RFP positive human supporting cells).11. Wash the cells one with 2 ml of PBS12. Add 2.0ml of fresh Endo-Growth medium (control) or Experimental media (EndoGrowth medium, plus pro- or anti-angiogenic regents according to customer’sneeds).13. Place the plate back into the incubator. Day 4, 6, 8, 10, 12, and 14……14. Replace the medium every 2 days until the end of the experiments.