Kruk P A, Maines-Bandiera S L, Auersperg N
Department of Anatomy, University of British Columbia, Vancouver, Canada.
Lab Invest. 1990 Jul;63(1):132-6.
The ovarian surface epithelium (OSE) is thought to give rise to over 85% of human ovarian carcinomas. In spite of its clinical importance, no animal models for the in vivo investigation of this tissue exist, and available culture methods have yielded limited success. In this study, OSE cells from 55 normal ovarian biopsy specimens were used to improve and simplify the methodology for OSE culture and to define the influence of clinical parameters on cultured OSE cells. An improved explanation method was developed which takes advantage of the tenuous attachment of OSE to underlying tissues: the surface epithelium was scraped off the ovarian surface with a rubber scraper, generating epithelial fragments which produced monolayers in culture, with little contamination by other cell types. The scrape method is superior to the explant method previously described (Siemens CH, Auersperg N: J Cell Physiol 134:347, 1988) in terms of speed, simplicity, higher purity of cultures, and increased cell yield. An improved nutrient medium (199/MCDB105/15%FBS) resulted in OSE lines that maintained the original epithelial phenotype for up to 12 population doublings. OSE, detached from the ovary, remained viable if frozen in liquid nitrogen either before culture or in primary culture on strips of plastic, providing OSE independently of the availability of surgical specimens. Growth was not influenced by diagnosis (nonmalignant gynecologic disorders), patient age (mean range: 40.5, 20 to 62 years), or the presence of inclusion cysts or large follicles in the biopsy specimen. This culture system provides conditions for in depth studies of OSE physiology and pathology.
卵巢表面上皮（OSE）被认为是超过85%的人类卵巢癌的起源。尽管其具有临床重要性，但目前尚无用于该组织体内研究的动物模型，现有的培养方法也成效有限。在本研究中，使用来自55例正常卵巢活检标本的OSE细胞来改进和简化OSE培养方法，并确定临床参数对培养的OSE细胞的影响。开发了一种改进的取材方法，该方法利用了OSE与下方组织的疏松附着：用橡胶刮刀从卵巢表面刮下表面上皮，产生上皮碎片，这些碎片在培养中形成单层，很少被其他细胞类型污染。在速度、简便性、培养物更高的纯度和增加的细胞产量方面，刮取法优于先前描述的外植体法（Siemens CH，Auersperg N：《细胞生理学杂志》134：347，1988）。一种改良的营养培养基（199/MCDB105/15%胎牛血清）使OSE细胞系在多达12次群体倍增中保持原始上皮表型。从卵巢分离的OSE，无论是在培养前还是在塑料条上的原代培养中液氮冷冻，均保持活力，从而可独立于手术标本的可用性提供OSE。生长不受诊断（非恶性妇科疾病）、患者年龄（平均范围：40.5岁，20至62岁）或活检标本中包涵囊肿或大卵泡的存在影响。该培养系统为深入研究OSE生理学和病理学提供了条件。
