Varani J, Perone P, Fligiel S E, Inman D R, Voorhees J J
Department of Pathology, University of Michigan Medical School, Ann Arbor 48109.
Arch Dermatol Res. 1994;286(8):443-7. doi: 10.1007/BF00371569.
Human dermal fibroblast and human epidermal keratinocyte survival was examined under various conditions in organ culture. Using cell recovery from organ-cultured tissue as the criterion, it was observed that no keratinocytes and few fibroblasts survived incubation for 10-12 days in serum-free basal medium containing a low level (0.15 mM) of extracellular Ca2+. Increasing the extracellular Ca2+ concentration to 1.4 mM or treating the tissue with 3 microM retinoic acid (RA) under low Ca2+ conditions resulted in increased keratinocyte and fibroblast survival; the two treatments together were more effective than either treatment alone. The same treatments preserved fibroblast survival when pieces of isolated dermal tissue were incubated in organ culture and also supported fibroblast survival in monolayer culture. These findings indicate that recovery of keratinocytes and fibroblasts from skin after maintenance in organ culture provides a simple but definitive measure of the viability of the major cellular elements present in the tissue. These findings suggest that RA treatment enhances survival of both fibroblasts and keratinocytes and that these effects of RA can be seen at physiological Ca2+ concentrations as well as at suboptimal levels of extracellular Ca2+. Finally, these results indicate that the dermis is a direct target of RA.
在器官培养的各种条件下检测了人皮肤成纤维细胞和人表皮角质形成细胞的存活情况。以从器官培养组织中回收细胞作为标准，观察到在含有低水平（0.15 mM）细胞外Ca2+的无血清基础培养基中孵育10 - 12天后，没有角质形成细胞存活，只有少数成纤维细胞存活。在低钙条件下，将细胞外Ca2+浓度提高到1.4 mM或用3 microM视黄酸（RA）处理组织，可增加角质形成细胞和成纤维细胞的存活；两种处理一起使用比单独任何一种处理都更有效。当将分离的真皮组织块在器官培养中孵育时，相同的处理可维持成纤维细胞的存活，并且在单层培养中也支持成纤维细胞的存活。这些发现表明，在器官培养中维持后从皮肤中回收角质形成细胞和成纤维细胞，为组织中主要细胞成分的活力提供了一种简单但明确的测量方法。这些发现表明，RA处理可提高成纤维细胞和角质形成细胞的存活率，并且在生理Ca2+浓度以及细胞外Ca2+的次优水平下都可以看到RA的这些作用。最后，这些结果表明真皮是RA的直接靶点。
