中国癌症防治杂志2025，Vol.17Issue(2)：150-156,7.DOI:10.3969/j.issn.1674-5671.2025.02.04
小鼠输卵管上皮类器官的培养与基因编辑
Cultivation and gene editing of mouse oviductal epithelial organoids
摘要
Abstract
Objective To establish oviductal epithelial organoids derived from Cas9-EGFP mice and perform gene editing.Methods Oviductal epithelial cells were isolated from Cas9-EGFP mice via enzymatic digestion and cultured in organoid-specific medium.The proliferation rates and size progression of organoids were monitored via continuous imaging.Cas9-EGFP expression was observed using fluorescence microscopy.Immunofluorescence staining was performed to confirm the origin of reproductive system and assess the expression of ciliated cell markers.Lentivirus carrying sgTrp53 was used to infect Cas9-EGFP mice oviductal epithelial organoids,with editing efficiency was validated by T7E1 assay.Results The abdominal cavity of mice was accessed through a midline incision,revealing coiled oviducts located between the uterine horns and ovaries.Oviductal epithelial organoids derived from Cas9-EGFP mice assembled into cystic structures by day 2 of culture,reaching diameters of 400 μm by day 7,with stable Cas9 protein expression.Immunofluores-cence confirmed their origin from the female murine reproductive system,and expression of ciliated cell markers.Following lentiviral transduction with sgTrp53,Trp53 mRNA expression was significantly reduced in edited organoids compared to unedited controls(P=0.021).Conclusions Oviductal epithelial organoids are successfully established from Cas9-EGFP mice,and CRISPR-Cas9-medi-ated gene editing was achieved in this model system.关键词
输卵管/类器官/基因编辑Key words
Oviducts/Organoids/Gene editing分类
医药卫生引用本文复制引用
杜一,岳静,陈婧瑶,周圣涛..小鼠输卵管上皮类器官的培养与基因编辑[J].中国癌症防治杂志,2025,17(2):150-156,7.基金项目
国家重点研发计划(2022YFA1106600) （2022YFA1106600）
