CAJ | 학술논문

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。
배경：과제조증성공적용 MV3흑소류세포화인각질형성세포체외중건흑소류삼유모형。 　　목적：이용MV3흑소류세포화HaCaT세포결합거표피적진피체외중건흑소류모형。 　　방법：MV3흑소류세포화HaCaT세포안조불동비례혼합접충우인거표피적진피조직상，채용액하배양화공기-액면배양상결합기술진행배양，체외구건조직공정피부모형。대소구건적피부흑소류모형진행상규절편면역조화관찰。 　　결과여결론：소목정-이홍염색현시MV3흑소류세포재거표피진피표면성층분포혹형성류조，HaCaT 세포화류세포혼합생장，형성전형적표피양결구。부분류세포침윤도거표피진피천층혹내부，성류조분포。CK10、CK-pan화S-100면역조화염색현시양성。수착MV3∶HaCaT세포비례적증고，CK10，CK-pan유표층축점하이，유층상분포변위단괴상분포，S-100단백염색칙분층축점명현，부분구역성류상분포。결과가견이용 MV3흑소류세포화 HaCaT 세포결합거표피진피체외가이구건피부흑소류모형。
BACKGROUND:We have built the three-dimensional human skin melanoma model with human epidermal keratinocytes and MV3 melanoma cel s co-cultured on the de-epidermized dermis in vitro. OBJECTIVE:To establish a skin model of melanoma by mixed culture of MV3 melanoma cel s and HaCaT cel s on the de-epidermized dermis in vitro. METHODS:MV3 melanoma cel s and HaCaT cel s were mixed with different percentages and inoculated on the surface of de-epidermized dermis fol owed by a liquid culture and air-liquid culture, and then the tissue-engineered skin model was established in vitro. Routine biopsy immunohistochemical observation was performed on the constructed skin melanoma model. RESULTS AND CONCLUSION:Hematoxylin-eosin staining showed that MV3 melanoma cel s were distributed on the surface layer of de-epidermized dermis and formed tumor masses, while the HaCaT cel s were mixed growth with tumor cel s and formed a typical epidermoid structure. Some tumor cel s infiltrated into the surface or deep of de-epidermized dermis and showed a tumor foci distribution. The CK10, CK-pan and S-100 proteins were positive for immunohistochemical staining. With the increasing of MV3:HaCaT cel percentage, CK10 and CK-pan gradual y down-moved from the surface, and changed from layer distribution to lumpy distribution, while the staining of S-100 protein was gradual y distributed layer-by-layer, and some area showed tumor-like distribution. The results show that the skin model of melanoma can be in vitro constructed successful y by mixed culture of MV3 melanoma cel s and HaCaT cel s on de-epidermized dermis.