携带HPV11型基因组细胞三维培养模型的建立及其衣壳蛋白L1的表达

目的用携带人乳头瘤病毒11型(HPV11)基因组的角质形成细胞(HPV11.HaCaT细胞),在体外建立皮肤类似培养物三维模型,观察HPV11衣壳蛋白L1是否表达,为进一步建立体外病毒复制系统和药效筛选模型奠定基础。方法用人成纤维细胞和I型鼠尾胶原制备真皮类似物凝胶,接种HPV11.HaCaT细胞进行三维培养。同时以正常HaCaT细胞作对照。15天时取皮肤类似物进行冰冻切片,HE染色进行组织结构检查,免疫组化法检查角蛋白10、泛角蛋白以及HPV11L1蛋白表达。结果HE染色显示,HPV11.HaCaT细胞和正常HaCaT细胞的层数明显增多。免疫组化显示,两种细胞的皮肤类似物角蛋白10、泛角蛋白表达全部阳性。HPV11.HaCaT细胞表达L1蛋白阳性,而正常HaCaT表达L1蛋白阴性。结论三维培养方式可诱导HPV11.HaCaT细胞分化,皮肤类似培养物出现细胞分化标志,并且表达HPV11型衣壳蛋白L1。

Establishment of three-dimensional culture model by HaCaT keratinocytes carrying HPV11 genome and expression of HPV 11 major capsid protein L1 in this model

Objective To study the expression of HPV 11 major capsid protein L1 in the skin equivalent model established by keratinocytes carrying HPV11 genome (HPV11.HaCaT cells) in vitro, which would serve as the basis for future studies of HPV replication system in vitro and anti-viral drugs screening model. Methods Cultured fibroblasts were mixed with rat tail type I collagen to form a dermal equivalent, HPV11.HaCaT cells were seeded on the surface of the dermal equivalent and cultured three-dimensionally. After 15...