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NCI-H292作为用于人类副粘病毒分离和增殖的替代细胞系。

NCI-H292 as an alternative cell line for the isolation and propagation of the human paramyxoviruses.

作者信息

Castells E, George V G, Hierholzer J C

机构信息

Respiratory and Enteric Viruses Branch, Centers for Disease Control, Atlanta, Georgia.

出版信息

Arch Virol. 1990;115(3-4):277-88. doi: 10.1007/BF01310536.

Abstract

Primary rhesus monkey kidney (MK) cells have long been the cells of choice for isolation and propagation of the human paramyxoviruses (parainfluenza 1, 2, 3, 4A, 4B, and mumps). However, problems with the supply and cost of MK cells and the presence of endogenous viruses, including herpes B virus and SV-5, necessitated a search for an alternative cell line. Continuous cell cultures of human origin (L132, A-549, HuT-292, HEK, G-293, G-401, A-498, A-704, CAKI-1, RD) and simian origin (LLC-MK2, BSC-1, MA-104, Vero) were evaluated for their capacity to support the growth of the human paramyxoviruses, as followed by cytopathic effect, hemadsorption, hemagglutination, and EIA. NCI-H292 (HuT-292) human lung mucoepidermoid carcinoma cells (ATCC # CRL-1848) proved to be the most sensitive line for cultivating all serotypes and strains of the paramyxoviruses. These cells were also shown to be a suitable substitute for MK in primary isolation of paramyxoviruses from clinical specimens. RPMI-1640 with 1.5 micrograms/ml trypsin was the preferred maintenance medium; alternatively, Eagle's MEM supplemented with 1.5 micrograms/ml trypsin and 0.1% ITS was satisfactory. NCI-H292 cells are a continuous line with excellent growth characteristics, although the genetic polyploidy of the cells may limit the number of passages of usable cells.

摘要

恒河猴原代肾细胞(MK细胞)长期以来一直是分离和培养人副粘病毒(副流感病毒1、2、3、4A、4B和腮腺炎病毒)的首选细胞。然而,MK细胞的供应和成本问题以及内源性病毒(包括B型疱疹病毒和SV - 5)的存在,使得人们需要寻找替代细胞系。对人源(L132、A - 549、HuT - 292、HEK、G - 293、G - 401、A - 498、A - 704、CAKI - 1、RD)和猴源(LLC - MK2、BSC - 1、MA - 104、Vero)的连续细胞培养物进行了评估,以确定它们支持人副粘病毒生长的能力,评估指标包括细胞病变效应、血细胞吸附、血凝和酶免疫分析。NCI - H292(HuT - 292)人肺黏液表皮样癌细胞(ATCC编号# CRL - 1848)被证明是培养所有血清型和毒株副粘病毒最敏感的细胞系。这些细胞也被证明是从临床标本中初次分离副粘病毒时MK细胞的合适替代品。含1.5微克/毫升胰蛋白酶的RPMI - 1640是首选的维持培养基;另外,补充有1.5微克/毫升胰蛋白酶和0.1% ITS的伊格尔氏MEM培养基也令人满意。NCI - H292细胞是具有优良生长特性的连续细胞系,尽管细胞的遗传多倍性可能会限制可用细胞的传代次数。

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