摘要
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： We have built the three-dimensional human skin melanoma model with human epidermal keraUnocytes and MV3 melanoma cells co-cultured on the de-epidermized dermis in vitro. OBJECTIVE： To establish a skin model of melanoma by mixed culture of MV3 melanoma calls and HaCaT cells on the de-epidermized dermis in vitro. METHODS： MV3 melanoma cells and HaCaT calls were mixed with different percentages and inoculated on the surface of de-epidermized dermis followed 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 calls were distdbuted on the surface layer of de-epidermized dermis and formed tumor masses, while the HaCaT cells were mixed growth with tumor cells and formed a typical epidermoid structure. Some tumor cells 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 cell percentage, CK10 and CK-pan gradually down-moved from the surface, and changed from layer distribution to lumpy distribution, while the staining of S-100 protein was gradually 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 successfully by mixed culture of MV3 melanoma calls and HaCaT calls on de-epidermized dermis.
出处
《中国组织工程研究》
CAS
CSCD
2013年第37期6620-6627,共8页
Chinese Journal of Tissue Engineering Research
基金
Guizhou Provincial Scientific and Technological Projects,No.D2009-7~~
