[Comparison of techniques irrigating and reversing rabbit ear vein to digest, isolate and culture endothelial cells].
OBJECTIVE
To establish the better method for isolating and culturing the endothelial cells (ECs).


METHODS
The "irrigative digestion" and "reverse interiorly-exteriorly digestion" methods were performed for digesting, isolating, collecting and culturing the ear vein endothelial cells of rabbit. The trypan blue stain was used to test the cell activity. The third-passage cell was identified by factor VI related antigen. The differences between the methods in cell number, activity and purity were compared to get an optimal method.


RESULTS
The number of ECs deriving from the "irrigative digestion" method had no significant difference from that deriving from the "reverse interiorly-exteriorly" method when cells were isolated from rabbit ear vein originally. However, after cultured for 5 or 10 days, the vein endothelial cells from the "irrigative digestion" method showed the growth status superior to those from the "reverse interiorly-exteriorly" method. The cultured cells had a cobble stone appearance with a strict monolayer growth, it could be observed under inverted microscope. The factor VIII related antigen was tested by immunohistochemistry, it supported that the cultured cells was ECs.


CONCLUSIONS
The "irrigative digestion" method is the better choice to isolate endothelial cells from small vessel.
Chen
Juan
J
The Department of Ophthalmology, the Red Cross Hospital of Yunnan Province, Kunming 650031, China.
Wang
Ting-hua
TH
Huang
Gui-qin
GQ
Yi
Wei-bin
WB
Xu
Yan-ze
YZ
Luo
Ze-wen
ZW
Hu
Zhu-lin
ZL
D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods*; D049109:Cell Proliferation; D002469:Cell Separation / Q000379:methods*; D004423:Ear / Q000098:blood supply*; D042783:Endothelial Cells / Q000166:cytology* / Q000378:metabolism; D005169:Factor VIII / Q000378:metabolism; D005260:Female; D007150:Immunohistochemistry; D008297:Male; D008853:Microscopy; D011817:Rabbits; D007507:Therapeutic Irrigation; D013997:Time Factors; D014680:Veins / Q000166:cytology
D005169:Factor VIII