Effects of epidermal growth factor on the proliferation and cell cycle regulation of cultured human amnion epithelial cells
Effects of epidermal growth factor on the proliferation and cell cycle regulation of cultured human amnion epithelial cells
复制标题
DOI：
10.1016/j.jbiosc.2012.03.021
复制
发表时间：
2012-08-01
影响因子：
2.8
通讯作者：
Hayati, Abdul Rahman
中科院分区：
文献类型：
--
作者：
Fatimah, Simat Siti;Tan, Geok Chin;Hayati, Abdul Rahman
人羊膜上皮细胞（HAECs）在再生医学的组织工程中具有巨大的应用前景。为此，需要大量的HAEC。因此，将外源性生长因子添加到培养基中以改善上皮细胞增殖。本研究的目的是确定表皮生长因子（EGF）对培养的HAECs的增殖和细胞周期调控的影响。将P1时的HAEC在含有HAM F12：Dulbecco改良Eagles培养基（1：1）的等体积混合物的培养基中培养7天，所述HAM F12：Dulbecco改良Eagles培养基补充有在还原血清中的不同浓度的EGF（0、5、10、20、30和50 ng/ml EGF）。形态学，生长动力学和细胞周期分析，流式细胞术进行了评估。还测定了细胞周期控制基因、多能转录因子、上皮基因和神经元基因的定量基因表达。EGF促进HAECs增殖的最适浓度为10 ng/ml。与对照组相比，EGF显著增加HAECs在细胞周期S期和G2/M期的比例。培养结束时，细胞周期分析显示HAECs仍为二倍体细胞。EGF可显著降低HAECs中p21、pRb、p53和GADD 45的mRNA表达。EGF还显著降低多能基因表达：Oct-3/4、Sox 2和Nanog;上皮基因表达：CK 14、p63、CK 1和Involucrin;神经元基因表达：NSE、NF-M和MAP 2。结果表明，EGF是一种强的促有丝分裂剂，通过细胞周期调节促进HAECs的增殖。EGF不能促进HAECs的分化和多能基因的表达。(c)2012年，生物技术学会，日本。All rights reserved.
Human amnion epithelial cells (HAECs) hold great promise in tissue engineering for regenerative medicine. Large numbers of HAECs are required for this purpose. Hence, exogenous growth factor is added to the culture medium to improve epithelial cells proliferation. The aim of the present study was to determine the effects of epidermal growth factor (EGF) on the proliferation and cell cycle regulation of cultured HAECs. HAECs at P1 were cultured for 7 days in medium containing an equal volume mix of HAM's F12: Dulbecco's Modified Eagles Medium (1:1) supplemented with different concentrations of EGF (0, 5, 10, 20, 30 and 50 ng/ml EGF) in reduced serum. Morphology, growth kinetics and cell cycle analysis using flow cytometry were assessed. Quantitative gene expression for cell cycle control genes, pluripotent transcription factors, epithelial genes and neuronal genes were also determined. EGF enhanced HAECs proliferation with optimal concentration at 10 ng/ml EGF. EGF significantly increased the proportion of HAECs at S- and G2/M-phase of the cell cycle compared to the control. At the end of culture, HAECs remained as diploid cells under cell cycle analysis. EGF significantly decreased the mRNA expression of p21, pRb, p53 and GADD45 in cultured HAECs. EGF also significantly decreased the pluripotent genes expression: Oct-3/4, Sox2 and Nanog; epithelial genes expression: CK14, p63, CK1 and Involucrin; and neuronal gene expression: NSE, NF-M and MAP 2. The results suggested that EGF is a strong mitogen that promotes the proliferation of HAECs through cell cycle regulation. EGF did not promote HAECs differentiation or pluripotent genes expression. (c) 2012, The Society for Biotechnology, Japan. All rights reserved.