3D培养细胞球快速石蜡切片方法建立及应用

Establishment and application of a rapid paraffin sectioning method for 3D cellular spheroids

目的建立一种适合于三维(3D)细胞培养模型的、简便快速的石蜡切片技术及其染色方法,为更便捷、快速地制作3D细胞切片并观察培养模型形态、功能,开展应用研究提供病理技术平台。方法利用超低吸附培养板将HepG2细胞或/和LX-2细胞培养成3D细胞球,制作石蜡切片,并进行HE染色、免疫组化染色和免疫荧光染色检测细胞球组织形态和蛋白表达。结果建立了适用于3D细胞培养模型的快速石蜡切片方法,制作的切片细胞球结构完整、细胞界限清楚,抗原活性良好,进行HE染色、免疫组化染色和免疫荧光染色能准确检测3D细胞球的形态结构,蛋白表达和分布。结论该快速石蜡切片方法的建立,丰富了3D细胞培养模型研究手段,可促进其在基础研究,转化医学和药物研发中的应用。

Objective To establish a simple and fast paraffin section technique and staining method suitable for use with three-dimensional(3 D)cell culture models and to provide a convenient pathological technology platform for the study of the morphology,function,and application of 3 D cell culture models.Methods HepG2 cells or/and LX-2 cells were cultured into 3 D multicellular spheroids using an ultra-low attachment surface culture plate,and paraffin sections were made.Hematoxylin and eosin(HE)staining,immunohistochemical staining,and immunofluorescence staining were performed to detect the histological and morphological characteristics and protein expression of the multicellular spheroids.Results A rapid paraffin sectioning method suitable for 3 D cell culture models was established.The prepared sectioned cell spheroids had complete structure,clear cell boundaries,and good antigen activity.The morphological structure and the expression and distribution of the proteins of 3 D cell culture models could be accurately detected by HE staining,immunohistochemical staining,and immunofluorescence staining.Conclusions The establishment of a rapid paraffin sectioning method has enriched the research method available for 3 D cell culture models and could promote their application in basic research,translational medicine,and drug development.