Home > Browse Issues > Vol.36 No.7
Isolation and Cultivation of Retinal Microvascular Pericytes from Weanling Rats
Yang Miqing1#, Liu Guanghui2#, Wang Hang1, Huang Chenwen3, Zheng Yongzheng2, Reng Bingyi2,Zhou Zixin2, Yang Wei1, Ji Binfeng1, Meng Chun1*
1College of Biological Science and Biotechnology, Fuzhou University, Fuzhou 350108, China;2Affiliated People’s Hospital, Fujian University of Traditional Chinese Medicine, Fuzhou 350004, China;3School of Life Science and Technology, Tongji University, Shanghai 200092, China
Abstract: The aim of this work was to culture retinal microvascular pericytes (RMPs) from weanling rat and optimize the protocol for isolation and cultivation of rat RMPs. The retinas were obtained separately from weanling rats (n=15) and adult rats (n=15) by ophthalmic microsurgery instruments. RMPs were isolated from the retinas by mechanical morcel， enzymatic digestion and filtration， and were cultivated. Cell growth was assessed by MTT. Morphological examination of RMPs was performed by inverted microscopy， and further characterization was analyzed by immunofluorescence. The differences in isolating-time， integrity-ratio， total-numbers， morphology， and marker-expression were observed. All retinas of weanling rats were successfully isolated. 24 of them were intact and 6 of them were broken fragmentarily. The time for isolating retina from a single eye was about 14.3~45.5 seconds. The isolating-time， integrity-ratio of 2 groups showed no significant difference (P>0.05). There was no significant difference in morphology and marker expression between the 2 groups. But the total numbers of primary RMPs and the proliferative ability of the cells from weanling rats were higher than those from adult rats. These results suggested that the retinas of weanling rats were favorable tissue source for RMPs culture. Here we established a protocol for the isolation and cultivation of RMPs from weanling rats.
CN
EN