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Published on: June 8, 2017
[A cell culture model in vitro for studying the permeability of cochlear microvascular endothelial cells in guinea
1Department of Otorhinolaryngology, Southwest Hospital, Third Military Medical University, Chongqing 400038, China. yingsc@public.cta.cq.cn
Objective:
To establish an "in vitro" system for studying the permeability of cochlear microvascular endothelial cells.
Methods:
Cochlear stria vascularis from guinea pigs were isolated by microsurgery, then cultured in vitro to get the monolayer cells of cochlear microvascular endothelium. The purity and property of cultured cells were identified by immunochemical method. The model in vitro was established by millcell technique of inoculation of the cultured monolayer cells of cochlear microvascular endothelium, then the permeability of this model was studied by 125I-bovine serum albumin. The positive group (brain microvascular endothelial cells), negative group (lung microvascular endothelial cells) and blank group (no cells) were conducted as control.
Results:
1. When cochlear stria vascularis fragment was isolated and cultured for two days, a few culture cells appeared around the fragments, then the amounts of culture cells increased. About ten days later, clusters of culture cells could be seen. Generally, spindle-shaped single culture cells were observed after cloning. When subcultured for eight days, there formed a monolayer tightly packed against the neighbouring cells; these cells showed the cobblestone shape under the microscope. For immunochemistry, over 95% of the cultured cells showed a positive reaction to factor VIII related antigen. 2. The permeability of 125I-bovine serum albumin in testing group was higher than that in the positive group (P < 0.05) and was lower than that in the negative group and the blank group (P < 0.01).
Conclusion:
An ideal model in vitro has been established for studying the permeability of cochlear microvascular endothelial cells at molecular level.

