Isolation and characterization of goat retinal microvascular endothelial cells.
In this report, we demonstrate a method for the isolation of pure homogeneous endothelial cell population from goat's eye without using multi-step procedure and sophisticated instrument facilities. Microvascular endothelial cell from goat's retina were isolated using enzymatic method and cultured in Iscove's Modified Dulbecco's Medium supplemented with 10% fetal bovine serum. Five to 7 d after plating, a monolayer of endothelial cells was formed. These cells were identified as endothelial cells by morphology and confirmed by positive immunocytochemistry for vWF, CD31, VE-cadherin, CD146, VCAM-1, and ICAM-1, a specific marker for endothelial cells. We have compared both the mechanical and non-mechanical enzymatic methods in isolating pure endothelial cells. Cells plated on 4% gelatin-coated dishes resulted in tubular morphology, a characteristic of endothelial cells. This method is simpler and cost-effective when compared with other previously reported methods. These endothelial cells will be more helpful to identify the role of various factors in angiogenic-related disease such as diabetic retinopathy.
Haribalaganesh
Ravinarayanan
R
Department of Biotechnology, Division of Molecular and Cellular Biology, Kalasalingam University (Kalasalingam Academy of Research and Education), Anand Nagar, Krishnankoil, 626 190, Tamilnadu, India.
Banumathi
Elayappan
E
Sheikpranbabu
Sardarpasha
S
Deepak
Venkataraman
V
Sirishkumar
Namagiri
N
Gurunathan
Sangiliyandi
S
D000818:Animals; D002469:Cell Separation / Q000379:methods*; D002478:Cells, Cultured; D042783:Endothelial Cells / Q000166:cytology* / Q000378:metabolism; D004730:Endothelium, Vascular / Q000166:cytology*; D007150:Immunohistochemistry; D012171:Retinal Vessels / Q000166:cytology* / Q000378:metabolism