Product Name 3D Human Pancreatic Microvascular
Catalog Number EP007
Product Format 6 , 12, and 24 well
Storage 37° C
Human pancreas development starts between 26 and 35 days post conception with the
emergence of dorsal and ventral buds from the foregut epithelium. At 6 weeks of gestation
(equivalent to 4 weeks post conception) the two buds fuse and become a single organ
formed by stratified epithelium embedded in mesenchyme. The stratified epithelium will
give rise to both the exocrine and endocrine compartments of the definitive pancreas.
One important physiological regulator of development and normal function of the
endocrine cells of the pancreas is the microcirculation through specialized sinusoidal
capillaries that irrigate the islets of Langerhans. The endothelial cells of these capillaries
are highly fenestrated to facilitate the exchange of signals. The dense network ensures that
each endocrine cell (glucagon-producing ?-cell, insulin-producing ?-cell, somatostatinproducing ?-cell, ghrelin-producing ?-cell and pancreatic polypeptide-producing PP-cell) is
in close proximity to the circulation. It makes up a considerable part of the islets and it is
responsible for critical communication via blood signals between the endocrine and
exocrine pancreas and also between the different cell types that populate the islets. After
transplantation of islets to the pancreas, angiogenesis is key to restoring proper function
Our 3D Human Pancreatic Microvascular Endothelial Angiogenesis model is construct
using GFP- Tagged human pancreatic microvascular endothelial cells are co-cultured with
RFP- Tagged human Supporting cells. GFP positive lymphatic capillary like tubule
formation can be monitored in real time under fluorescence microscope throughout the
whole process of the experiment.
1) Cells used in the 3D model are all human cells; results obtained are more relevant to
human situations rather than those data from animal models, i.e. CAM et al.
2) The whole angiogenesis process can be monitored (from cell inoculation to the end
of experiment), therefore, more crucial information can be acquired at multiple time
points from a single experiment.
3) No need to perform post-experimental staining for endothelial markers, this is
particularly important, if those markers are changed in experimental conditions
involved in the studies.
The 3D Human Pancreatic Microvascular Endothelial Angiogenesis contains all of the
materials necessary to perform multiple angiogenesis assays in 6, 12, or 24 well formats.
The 3D model is designed that the testing materials, i.e. compounds, conditioned media, or
tissue explants, can be added into the system at any time, ranging from the onset of
vasculogenesis to advanced angiogenesis. The resulting effect on tubule formation (tubular
length, number of branches et al) can be monitored throughout the whole process under
inverted fluorescence microscope.
Reagents and Materials Provided:
(1) 1 x vial of mixture of Human Pancreatic Microvascular Endothelial Angiogenesis ECs
and RFP-tagged supporting cells (-80°C or liquid N2)
(2) 1 x 24-well Alpha Coat Solution coated plate (Room temperature, for 2 months)
(3) 1 x 500ml of Endo-Growth Medium (4°C)
Protocols: Day 1
1. Pre warm Endo-Growth Medium to 37ºC in a water bath
2. 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).
9. Take the plate from the incubator and examine cells under inverted fluorescence
microscopy (GFP Human Pancreatic Microvascular Endothelial Angiogenesis should
sparsely and evenly distributed among RFP positive human supporting cells).
10. Wash the cells one with 2 ml of PBS
11. Add 2.0ml of fresh Endo-Growth medium (control) or Experimental media (Endo-Growth
medium, plus pro- or anti-angiogenic regents according to customer’s needs).
12. Place the plate back into the incubator. Day 4, 6, 8, 10, 12, and 14……
13. Replace the medium every 2 days until the end of the experiments.