Department of Dermatology, Leiden University Medical Center, Room T-02-09, Postal Zone S2-P, P.O. Box 9600, 2300 RC, Leiden, The Netherlands.
Biogerontology. 2013 Apr;14(2):131-40. doi: 10.1007/s10522-013-9416-9. Epub 2013 Mar 16.
Serial passaging has a profound effect on primary cells. Since serially passaged cells show signs of cellular aging, serial passaging is used as an in vitro model of aging. To relate the effect of in vitro aging more to in vivo aging, we generated human skin equivalents (HSEs). We investigated if HSEs generated with late passage fibroblasts show characteristics of aged skin when compared with HSEs generated with early passage fibroblasts. Late passage fibroblasts had enlarged cell bodies and were more often positive for myofibroblast marker α-smooth muscle actin, senescence associated β-galactosidase and p16 compared with early passage fibroblasts. Skin equivalents generated with late passage fibroblasts had a thinner dermis, which could partly be explained by increased matrix metalloproteinase-1 secretion. In equivalents generated with late passage fibroblasts epidermal expression of keratin 6 was increased, and of keratin 10 slightly decreased. However, epidermal proliferation, epidermal thickness and basement membrane formation were not affected. In conclusion, compared with HSEs generated with early passage fibroblasts, HSEs generated with late passage fibroblasts showed changes in the dermis, but no or minimal changes in the basement membrane and the epidermis.
传代对原代细胞有深远的影响。由于连续传代的细胞表现出细胞衰老的迹象，因此连续传代被用作衰老的体外模型。为了使体外衰老更能与体内衰老相关，我们生成了人皮肤等效物（HSEs）。我们研究了与早期传代成纤维细胞相比，用晚期传代成纤维细胞生成的 HSE 是否表现出老化皮肤的特征。与早期传代成纤维细胞相比，晚期传代成纤维细胞的细胞体增大，并且更常呈肌成纤维细胞标志物α-平滑肌肌动蛋白、衰老相关β-半乳糖苷酶和 p16 阳性。用晚期传代成纤维细胞生成的皮肤等效物真皮较薄，这部分可以通过增加基质金属蛋白酶-1 的分泌来解释。在用晚期传代成纤维细胞生成的等效物中，角蛋白 6 的表皮表达增加，角蛋白 10 的表皮表达略有减少。然而，表皮增殖、表皮厚度和基底膜形成不受影响。总之，与用早期传代成纤维细胞生成的 HSEs 相比，用晚期传代成纤维细胞生成的 HSEs 在真皮中发生了变化，但基底膜和表皮没有或几乎没有变化。
