A modified medium that significantly improves the growth of human normal ovarian surface epithelial (OSE) cells in vitro
A modified medium that significantly improves the growth of human normal ovarian surface epithelial (OSE) cells in vitro
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DOI：
10.1038/labinvest.3700093
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发表时间：
2004-07-01
影响因子：
5
通讯作者：
Gayther, SA
中科院分区：
文献类型：
--
作者：
Li, NFF;Wilbanks, G;Gayther, SA
大约90%的恶性卵巢肿瘤是上皮性的，被认为是由单细胞层，即卵巢表面上皮细胞产生的。在培养中，人正常卵巢表面上皮（OSE）细胞在衰老前的寿命非常有限，很少超过10个群体的生存期。这限制了正常OSE细胞用于研究卵巢表面上皮生物学和鉴定导致恶性转化的分子事件。我们研究了使用改良培养基体外培养人正常OSE细胞的条件。在补充有表皮生长因子、氢化可的松、胰岛素和牛垂体提取物的改良培养基（NOSE-CM）中培养正常OSE细胞，与在基本的、未补充的培养基（III和先前使用的培养基（F-12 K培养基和William培养基E）中培养相比，导致接种和克隆效率、总体细胞生长和寿命的显著改善。在NOSE-CM中培养的细胞平均经历19.0个群体倍增（95%CI 16.3-21.7）;在BM中培养的细胞在类似的时间段内经历0.43-3.52个群体倍增。不同细胞系的生长曲线表明，OSE细胞在NOSE-CM中继续生长超过第11代，直至第18代，但在BM中培养时从未超过第7代。很可能建立OSE细胞体外生长的最佳条件将使上皮性卵巢癌转化的生物学和遗传学机制的研究成为可能。
Approximately 90% of malignant ovarian tumours are epithelial and thought to arise from a single cell layer, the ovarian surface epithelium. In culture, human normal ovarian surface epithelial (OSE) cells have a very limited lifespan before they senesce, rarely progressing beyond 10 population doublings. This has restricted the use of normal OSE cells for studying the biology of ovarian surface epithelium and identifying molecular events that contribute to malignant transformation. We have investigated the conditions for culturing human, normal OSE cells in vitro using modified media. Culturing normal OSE cells in a modified medium (NOSE-CM) supplemented with epidermal growth factor, hydrocortisone, insulin and bovine pituitary extract led to significant improvements in the seeding and cloning efficiencies, overall cell growth and lifespan compared to culturing in a basic, nonsupplemented medium (Ill and previously used media (F-12K medium and William's medium E). Cells cultured in NOSE-CM underwent, on an average, 19.0 population doublings (95% CI 16.3-21.7); cells cultured in BM underwent 0.43-3.52 population doublings over a similar time period. Growth curves established for different lines indicated that OSE cells continued to grow beyond passage 11 and up to passage 18 in NOSE-CM, but never beyond passage 7 when cultured in BM. It is likely that establishing optimal conditions for the growth of OSE cells in vitro will enable studies of the biological and genetic mechanisms of transformation in epithelial ovarian cancers.