In vitro cultivation and identification of retinal photoreceptor precursor cells from newborn mice / 解放军医学杂志

ABSTRACT

Objective To isolate and identify the photoreceptor precursor cells isolated from newborn mouse retina. Methods Fourteen newborn littermate mice were randomly divided into seven groups (P1-P7,2 each). The retinal cells were isolated from the newborn mice on postnatal 1-7d respectively,and then cultured in vitro. The cellular growth state was observed under inverted phase contrast microscope. After in vitro expansion,5-bromo-2-deoxy-uridine (BrdU),neuroepithelial stem protein (Nestin),neural retina leucine zipper (Nrl) and Opsin in cultured cells were detected by immunohistochemistry and immunofluorescence techniques. Results Most of retinal cells isolated from the seven groups (P1-P7) of newborn mice proliferated and adhered to the plate in the particular medium. Most of the growing cells were BrdU-positive cells. A part of the cells could express neural stem cell specific antigen-Nestin,the positive expression rate of Nestin in the seven groups (P1-P7) were 31.0%,31.6%,32.3%,30.2%,31.2%,30.9% and 29.5%,respectively,with no significant difference. Some growing cells expressed Nrl,and the expression of Nrl was increased significantly on the third day after birth,and then increased in small range,the positive expression rate of Nrl in the seven groups (P1-P7) were 20.6%,35.2%,65.5%,68.6%,71.6%,73.0% and 73.3%,respectively. The positive expression of Opsin was found only in a few cells of P5-P7,while the cells in groups P1-P4 did not express Opsin. Conclusions The photoreceptor precursor cells are presented in neonatal mouse retina. They have the ability of proliferation and may differentiate into retinal photoreceptor cells,having a potentiality of expressing Opsin. The photoreceptor precursor cells are increased significantly on the third day after birth,and then differentiate into mature retinal photoreceptor cells gradually.