The in vitro expression and secretion of vascular endothelial growth factor from free and alginate-polyornithine encapsulated choroid plexus epithelium.
The choroid plexus (CP) is a transplantable cell source secreting tropic and trophic factors for the treatment of brain and peripheral trauma characterized by cellular loss or dysfunction. Here we characterize the expression and secretion of vascular endothelial growth factor (VEGF) from neonatal porcine CP. Light and electron microscopy revealed that enzymatic digestion of the CP produced a preparation consisting primarily of epithelial cells without notable contaminating cells. Microarray analysis, quantitative polymerase chain reaction, and enzyme-linked immunosorbent assay were used to quantify the nuclear, cytoplasmic, and secretory compartmentalization of VEGF. In vitro, the kinetics of VEGF release were orderly, with stepwise increases in secretion over time. The secretory profile of VEGF from CP grown in configurations ranging from a simple monolayer to free-floating 3-dimensional clusters to clusters encapsulated within alginate-polyornithine microcapsules was similar. VEGF output was not affected notably when the cells were maintained in 90% stress medium or in other maintenance media devoid of serum proteins. Secreted VEGF was bioactive, as confirmed by demonstrating its continued ability to proliferate co-cultured human umbilical vascular endothelial cells. The robust ability of these cells to continue to secrete VEGF (and presumably other bioactive proteins) across a variety of dimensional configurations and medium types has implications for their use in clinical indications requiring novel and imaginative use of engineered ectopic transplant sites.
Thanos
Chris G
CG
LCT Biopharma, Inc., Providence, Rhode Island, USA. cgthanos@gmail.com
Schneider
Patricia A
PA
Bintz
Briannan E
BE
Jensen
Roderick
R
Bryant
Bronwyn
B
Bell
William J
WJ
Hudak
Jebecka
J
Emerich
Dwaine F
DF
D000464:Alginates / Q000737:chemistry; D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods; D002478:Cells, Cultured; D002831:Choroid Plexus / Q000166:cytology* / Q000378:metabolism*; D020099:Coated Materials, Biocompatible / Q000737:chemistry; D004847:Epithelial Cells / Q000166:cytology* / Q000378:metabolism*; D020723:Glucuronic Acid / Q000737:chemistry; D006603:Hexuronic Acids / Q000737:chemistry; D008422:Materials Testing; D010455:Peptides / Q000737:chemistry*; D013552:Swine; D023822:Tissue Engineering / Q000379:methods*; D042461:Vascular Endothelial Growth Factor A / Q000378:metabolism*
D000464:Alginates; D020099:Coated Materials, Biocompatible; D006603:Hexuronic Acids; D010455:Peptides; D042461:Vascular Endothelial Growth Factor A; C008973:polyornithine; D020723:Glucuronic Acid