ROCK inhibitor Y-27632 promotes proliferation of human keratinocytes and inhibits its senescence

doi:10.3969/j.issn.2095-4344.0800

Abstract

Abstract:

BACKGROUND: Facilitating the proliferation of keratinocytes in vitro is of great significance for the studies on dermatobiology and the pathogenesis of skin-related diseases.
OBJECTIVE: To investigate the effects of Rho kinase inhibitor Y-27632 on the proliferation and senescence of keratinocytes.
METHODS: Human keratinocytes were successfully isolated and the potential effect of Y-27632 (10 μmol/L) on the human keratinocytemorphology was investigated. The effect of Y-27632 on the proliferation of keratinocytes was investigated by population doubling curve; β-galactosidase staining was performed at P0 and P5 cells respectively to show the effect of Y-27632 on the senescence of keratinocytes. Finally, the proliferation of keratinocytes was detected by MTT assay and BrdU kit simultaneously.
RESULTS AND CONCLUSION: Human keratinocytes were of cobble-stone morphology and equal size, which kept great proliferation capacity at early stage. With the continuous passage of human keratinocytes, their proliferation capacity was declined gradually, showing a flat morphology and an enlarged cytoplasmic volume. The population doubling of human keratinocytes was increased significantly in the presence of Y-27632, and according to the β-galactosidase staining results, Y-27632 decreased the number of positive cells. Furthermore, the results of MTT assay and BrdU kit indicated that Y-27632 facilitated the proliferation of keratinocytes obviously in vitro. That is to say, when the human keratinocytes are cultured in the presence of Y-27632, the senescence of keratinocytes is inhibited. Meanwhile, the proliferation of keratinocytes is significantly increased, and our data shed new insight on the in vitro proliferation of human keratinocytes in future studies.

中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程

Key words: Tissue Engineering, Cell Proliferation, Burns

CLC Number:

R318

Yang Jin1, 2, Xiong Xia1. ROCK inhibitor Y-27632 promotes proliferation of human keratinocytes and inhibits its senescence[J]. Chinese Journal of Tissue Engineering Research, 2018, 22(24): 3869-3874.

Figures/Tables 1

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