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背景:课题组曾成功的用MV3黑素瘤细胞和人角质形成细胞体外重建黑素瘤三维模型。目的:利用MV3黑素瘤细胞和HaCaT细胞结合去表皮的真皮体外重建黑素瘤模型。方法:MV3黑素瘤细胞和HaCaT细胞按照不同比例混合接种于人去表皮的真皮组织上,采用液下培养和空气-液面培养相结合技术进行培养,体外构建组织工程皮肤模型。对所构建的皮肤黑素瘤模型进行常规切片免疫组化观察。结果与结论:苏木精-伊红染色显示MV3黑素瘤细胞在去表皮真皮表面成层分布或形成瘤灶,HaCaT细胞和瘤细胞混合生长,形成典型的表皮样结构。部分瘤细胞浸润到去表皮真皮浅层或内部,成瘤灶分布。CK10、CK-pan和S-100免疫组化染色显示阳性。随着MV3∶HaCaT细胞比例的增高,CK10,CK-pan由表层逐渐下移,由层状分布变为团块状分布,S-100蛋白染色则分层逐渐明显,部分区域成瘤状分布。结果可见利用MV3黑素瘤细胞和HaCaT细胞结合去表皮真皮体外可以构建皮肤黑素瘤模型。
Background: The research team successfully reconstructed melanoma three-dimensional model with MV3 melanoma cells and human keratinocytes in vitro. OBJECTIVE: To reconstruct the melanoma model by using MV3 melanoma cells and HaCaT cells in combination with dendritic dermis. Methods: MV3 melanoma cells and HaCaT cells were inoculated into dermis of human epidermis in different proportions and cultured. The tissue culture skin model was constructed in vitro by using the combination of submerged culture and air-liquid culture. The constructed skin melanoma model was routinely sectioned immunohistochemically. RESULTS AND CONCLUSION: The hematoxylin-eosin staining showed that the MV3 melanoma cells were stratified or formed neoplastic lesions on the surface of de dermal dermis, and HaCaT cells and tumor cells mixed to form a typical epidermal-like structure. Some of the tumor cells infiltrated into the superficial epidermis dermis or internal, tumor-forming distribution. CK10, CK-pan and S-100 immunohistochemical staining showed positive. With the increase of the ratio of MV3: HaCaT cells, CK10 and CK-pan gradually decreased from the surface to the clumps, while the S-100 protein stained gradually became more stratified and some areas became tumorous. The results show that using MV3 melanoma cells and HaCaT cells in combination with dendritic dermis in vitro skin melanoma model can be constructed.