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建立并鉴定麦哲伦拟海鞘胚胎细胞系。

Establishment and characterization of an embryonic cell line from Sarconesiopsis magellanica.

机构信息

Laboratorio de Entomología Médica y Forense, Escuela de Medicina y Ciencias de La Salud, Universidad Del Rosario, Calle 63D 24-31, Bogotá, Colombia.

出版信息

J Insect Sci. 2013;13:130. doi: 10.1673/031.013.13001.

Abstract

Sarconesiopsis magellanica (Le Guillou) (Diptera: Calliphoridae) is a necrophagous fly that is important in both human and veterinary medicines. This insect has been registered in Colombia as a biological indicator in estimating post-mortem interval. Insect cell cultures are an important biotechnological tool for basic and applied studies, and cell cultures derived from S. magellanica embryonic tissues are described in this study. S. magellanica embryonated eggs were taken for tissue explants. These were seeded in L-15, Grace/L-15, Eagle MEM, MM, VP12, MM/VP12, and Schneider culture media. The morphological, cytogenetic, biochemical, and molecular characteristics of the cell cultures were examined. Cell growth was achieved in the L15, Grace/L15, and Schneider culture media, and the confluent monolayers were obtained 8, 10, and 19 days after the embryonated eggs were explanted. However, the Schneider medium was the most efficient to develop the subcultures, and 21 passages have been maintained. The cell morphology of the primary cell cultures was initially heterogeneous, but in the confluent monolayer and in the subcultures there was greater cell morphology uniformity, fibroblastoid types being predominant. Cultured cells had a chromosomal number of 12, and the karyotypic complement consisted of five pairs of somatic chromosomes and one sexual pair. The cell culture isozyme patterns of S. magellanica coincided with adult samples from the same species. The molecular analysis, using RAPD-PCR, demonstrated the authentication of the cell cultures of this fly and their differentiation from other cultures derived from two sand flies species. This cell line is a new in vitro model that will be used in biomedical and biotechnological studies.

摘要

麦加利长足虻(Sarconesiopsis magellanica)(双翅目:丽蝇科)是一种重要的腐生性昆虫,在人类和兽医医学中都有应用。这种昆虫已在哥伦比亚被注册为估计死后间隔时间的生物指标。昆虫细胞培养是基础和应用研究的重要生物技术工具,本研究描述了从麦加利长足虻胚胎组织中获得的细胞培养。从麦加利长足虻受精卵中取出组织外植体。将这些组织外植体接种于 L-15、Grace/L-15、Eagle MEM、MM、VP12、MM/VP12 和 Schneider 培养基中。对细胞培养的形态学、细胞遗传学、生物化学和分子特征进行了研究。在 L15、Grace/L15 和 Schneider 培养基中实现了细胞生长,在外植后的第 8、10 和 19 天获得了融合的单层。然而,Schneider 培养基最有利于开发亚培养物,已经维持了 21 代。原代细胞培养的细胞形态最初是异质的,但在融合的单层和亚培养物中,细胞形态的均匀性更高,成纤维细胞类型占主导地位。培养细胞的染色体数为 12,核型组成由五对体细胞染色体和一对性染色体组成。麦加利长足虻细胞培养同工酶模式与来自同一物种的成虫样本一致。使用 RAPD-PCR 的分子分析证明了该蝇细胞培养的真实性及其与来自两种沙蝇物种的其他培养物的分化。这种细胞系是一种新的体外模型,将用于生物医学和生物技术研究。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2288/4014039/08304eeb76d5/f01_01.jpg

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