Reconstitution of the histologic characteristics of a giant congenital nevomelanocytic nevus employing the athymic mouse and a cultured skin substitute.
Reconstitution of the histologic characteristics of a giant congenital nevomelanocytic nevus employing the athymic mouse and a cultured skin substitute.
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使用无胸腺小鼠和培养的皮肤替代品重建巨大先天性色素痣的组织学特征。
DOI：
10.1111/1523-1747.ep12483707
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发表时间：
1991
期刊：
影响因子：
--
通讯作者：
Frank,DH
中科院分区：
文献类型：
--
作者：
Cooper,ML;Spielvogel,RL;Hansbrough,JF;Boyce,ST;Frank,DH
本研究旨在建立人类巨大先天性痣黑素细胞痣（GCNN）动物模型。皮肤移植物由1）来自12个月大GCNN患者的非受累断层皮肤，2）来自同一GCNN患者的痣断层皮肤，3）痣全层皮肤和4）尸体人断层皮肤制成。对于组1）和2），酶促分离人表皮和真皮细胞并在组织培养物中扩增。通过将培养的真皮细胞置于胶原-糖胺聚糖（GAG）基质中，然后将表皮细胞置于基质的相对层压侧，制备复合移植物。将所有移植物置于无胸腺小鼠的全层伤口上，并在6至38周后获得活组织切片用于光学显微镜（包括S-100免疫过氧化物酶染色）和电子显微镜。第1组小鼠均未出现肉眼或组织学病变。所有的痣全厚度小鼠保留痣大体。第2组黑色隆起斑块38周时的组织学检查显示与第3组相似的重建真皮。痣细胞更大，更多的上皮样在上层真皮，看到真正的GCNN。在24周时，在真皮中未观察到这些痣黑素细胞，这表明在此时间范围内（24-38周），痣细胞从培养移植物的表皮成分迁移到真皮。用S-100免疫过氧化物酶染色和电子显微镜证实了这些细胞的黑素细胞身份。这些发现是这种复合培养移植系统所特有的。培养特定类型的黑素细胞并将其置于无胸腺小鼠的皮肤替代物中的能力为GCNN和体内黑素细胞生物学的研究提供了基础。
This study addresses the development of an animal model for human giant congenital nevomelanocytic nevi (GCNN). Skin grafts were made from 1) non-involved spilt-thickness skin from a 12-month-old GCNN patient, 2) nevus split-thickness skin from the same GCNN patient, 3) nevus full-thickness skin, and 4) cadaveric human split-thickness skin. For groups 1) and 2), human epidermal and dermal cells were enzymatically isolated and expanded in tissue culture. Composite grafts were made by placing the cultured dermal cells into a collagen-glycosaminoglycan (GAG) matrix, followed by placement of the epidermal cells onto the opposite, laminated side of the matrix. All grafts were placed onto full-thickness wounds of athymic mice and biopsies were obtained from 6 to 38 weeks later for light microscopy including S-100 immunoperoxidase staining, and electron microscopy.The GCNN cultured skin mice (group 2) developed black, raised skin in the healed wounds. None of the group 1 mice developed lesions, grossly or histologically. All of the nevus full-thickness mice retained the nevus grossly. Histopathologic examination at 38 weeks of the black, raised plaques of group 2 demonstrated a reconstituted dermis similar to group 3. Nevus cells were larger and more epithelioid in the upper dermis, as seen with true GCNN. These nevomelanocytes were not seen in the dermis at 24 weeks, suggesting that the nevus cells migrated from the epidermal component of the cultured graft to the dermis during this time frame (24-38 weeks). The melanocyte identity of these cells was confirmed with S-100 immunoperoxidase staining and electron microscopy.These findings are unique to this composite cultured graft system. The ability to culture specific types of melanocytes and place them into skin substitutes on athymic mice provides a basis for the study of GCNN and melanocyte biology in vivo.