Krikun Graciela, Mor Gil, Alvero Ayesha, Guller Seth, Schatz Frederick, Sapi Eva, Rahman Mizanur, Caze Rebeca, Qumsiyeh Mazin, Lockwood Charles J
Department of Obstetrics, Gynecology and Reproductive Sciences, Yale University School of Medicine, 333 Cedar Street, P.O. Box 208063, New Haven, Connecticut 06520-8063, USA.
Endocrinology. 2004 May;145(5):2291-6. doi: 10.1210/en.2003-1606. Epub 2004 Jan 15.
Obtaining primary human endometrial stromal cells (HESCs) for in vitro studies is limited by the scarcity of adequate human material and the inability to passage these cells in culture for long periods. Immortalization of these cells would greatly facilitate studies; however, the process of immortalization often results in abnormal karyotypes and aberrant functional characteristics. To meet this need, we have introduced telomerase into cultured HESCs to prevent the normal shortening of telomeres observed in adult somatic cells during mitosis. We have now developed and analyzed a newly immortalized HESC line that contains no clonal chromosomal structural or numerical abnormalities. In addition, when compared with the primary unpassaged parent cells, the new cell line displayed similar biochemical endpoints after treatment with ovarian steroids. Classical decidualization response to estradiol plus medroxyprogesterone acetate were seen in both morphologically, and progestin was seen to induce or regulate the expression of IGF binding protein-1, fibronectin, prolactin, tissue factor, plasminogen activator inhibitor-1, and Fas/Fas ligand. In summary, an immortalized HESC line has been developed that is karyotypically, morphologically, and phenotypically similar to the primary parent cells, and it is a powerful and consistent resource for in vitro work.
获取原代人子宫内膜基质细胞(HESC)用于体外研究受到充足人类材料稀缺以及无法在培养中长时间传代这些细胞的限制。使这些细胞永生化将极大地促进研究;然而,永生化过程往往会导致异常的核型和异常的功能特征。为满足这一需求,我们已将端粒酶导入培养的HESC中,以防止在有丝分裂过程中成年体细胞中观察到的端粒正常缩短。我们现已开发并分析了一种新的永生化HESC系,该系不存在克隆性染色体结构或数量异常。此外,与未传代的原代亲本细胞相比,新细胞系在用卵巢类固醇处理后显示出相似的生化终点。在形态学上均观察到对雌二醇加醋酸甲羟孕酮的经典蜕膜化反应,并且发现孕激素可诱导或调节胰岛素样生长因子结合蛋白-1、纤连蛋白、催乳素、组织因子、纤溶酶原激活物抑制剂-1以及Fas/Fas配体的表达。总之,已开发出一种永生化HESC系,其核型、形态和表型与原代亲本细胞相似,并且是体外研究的强大且一致的资源。