Sorrell J M, Baber M A, Caplan A I
Skeletal Research Center, Department of Biology, Case Western Reserve University, Cleveland, Ohio 44106.
Tissue Eng. 1996 Spring;2(1):39-49. doi: 10.1089/ten.1996.2.39.
Bilayered dermal equivalents were constructed by seeding human papillary and reticular dermal fibroblasts into separate layers of type I collagen and allowing these layers to gel into a single entity. That these bilayered gels had fused was established through histologic examination and from the fact that these gels, when detached, contracted as a single unit. Papillary and reticular dermal fibroblasts remained in their respective layers as established by differentially labeling these dermal cells with fluorescent vital dyes l,l'-dioctacecyl-3,3,3',3'-tetramethylindocabocyanine perchlorate, DiIC(18)(3) (Dil), and 3,3'-dioctadecyloxacarbocyanine perchlorate, DiOC(18)(3) (DiO). The labeling with these vital dyes did not interfere with the ability of the cells to proliferate or to contract floating type I collagen gels. Thus, these bilayered gels can provide the means of creating dermal equivalents that contain a variety of different dermal cell types to assess their relative abilities, either alone or in various combinations, to support keratinocyte proliferation and differentiation and to contribute, eventually, to the formation of a multilayered skin equivalent.
通过将人乳头层和网状层真皮成纤维细胞接种到I型胶原蛋白的不同层中，并使这些层凝胶化形成一个单一实体，构建了双层真皮替代物。通过组织学检查以及这些凝胶在分离时作为一个单一单元收缩这一事实，证实了这些双层凝胶已经融合。通过用荧光活性染料1,1'-二辛基-3,3,3',3'-四甲基吲哚羰花青高氯酸盐（DiIC(18)(3)，Dil）和3,3'-二辛基氧杂羰花青高氯酸盐（DiOC(18)(3)，DiO）对这些真皮细胞进行差异标记，确定乳头层和网状层真皮成纤维细胞仍保留在各自的层中。用这些活性染料标记并不干扰细胞增殖或收缩漂浮的I型胶原蛋白凝胶的能力。因此，这些双层凝胶可以提供一种创建真皮替代物的方法，该替代物包含多种不同的真皮细胞类型，以评估它们单独或各种组合支持角质形成细胞增殖和分化以及最终促进形成多层皮肤替代物的相对能力。
