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Updated: Aug 29, 2026

Generation and Culturing of Primary Human Keratinocytes from Adult Skin
Published on: December 22, 2017
[Modulation of PKC inhibitor on growth cycle of human keratocyte]
1Ophthalmic Center of Peking University, the First Hospital, Peking University, Beijing 100034, China. yxm23@hotmail.com
Objective:
To study the effects of staurosporine (ST), an inhibitor of protein kinase C (PKC), on the cell cycle of cultured human keratocyte at G(1)/S phase transition and to study the molecular mechanism of this process.
Methods:
Flow cytometry technique was used to define the effect of ST on the cell cycle of cultured human keratocyte. Cyclin E protein levels of corneal keratocyte were analyzed by using the cyclin/DNA two parameter flow cytometry.
Results:
Treatment of ST (2.5 microg/L and 5 microg/L) for 18 hours blocked the growth of keratocyte in G(1) phase; the percentage of cells staying in G(1) phase was increased from 72.4% in the control group to 78.4% (group 2.5 microg/L ST) and 90.9% (group 5 microg/L ST), respectively. This effect was dose dependent. Cyclin E protein level in G(1) phase was decreased from 75.6% in the control group to 62.0% (group 2.5 microg/L ST) and 51.1% (group 5 microg/L ST) in cells treated with ST.
Conclusions:
PKC activity is important in promoting keratocyte proliferation. Inhibition of PKC activity prohibits G(1)/S phase progress and reduces cyclin E protein level. Inhibitor of PKC is involved in the modulation of the cell cycle of human keratocyte and may play an important role in inhibiting cell growth in G(1)/S phase.
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