A simple method for the in vitro culture of human retinal capillary endothelial cells.
A simple method for the culture of human retinal capillary endothelial cells in vitro is described in this report. Through a process of gentle mechanical disruption and sieving, a sufficient yield of retinal microvessels was obtained from one or two human eyes to allow the culture of endothelial cells in abundance. The cells were factor VIII-positive and demonstrated typical vascular endothelial morphology. Pericyte contamination was prevented by using human platelet-poor serum in primary culture and passaging cells with a low concentration of trypsin. Proliferation assays revealed that the cells grew best in Iscove's modified Dulbecco's modified Eagle's medium with 15% fetal calf serum (FCS). There was no difference in induced proliferation when FCS was compared to human platelet-poor serum. The method seemed to be as good as and much simpler than other recently described techniques. The study of human retinal capillary endothelial cells cultured in this way may shed light on the earliest stages of diabetic retinopathy and other diseases of the retinal microvasculature, particularly AIDS-related retinopathy and radiation retinopathy.
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Department of Ophthalmology, Prince of Wales Hospital, Randwick, NSW, Australia.
Gillies
M C
MC
D002196:Capillaries / Q000166:cytology / Q000378:metabolism; D002455:Cell Division; D002478:Cells, Cultured; D003470:Culture Media; D046508:Culture Techniques / Q000379:methods; D004730:Endothelium, Vascular / Q000166:cytology* / Q000378:metabolism; D005169:Factor VIII / Q000378:metabolism; D005455:Fluorescent Antibody Technique; D006801:Humans; D012171:Retinal Vessels / Q000166:cytology* / Q000378:metabolism; D014471:Umbilical Veins / Q000166:cytology / Q000378:metabolism
D003470:Culture Media; D005169:Factor VIII