Establishment and identification of a new cell line from<em> Culex pipiens quinquefasciatus</em> (Diptera: Culicidae)
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ACTA ENTOMOLOGICA SINICA  2018, Vol. 61 Issue (1): 79-85    DOI: 10.16380/j.kcxb.2018.01.009
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Establishment and identification of a new cell line from Culex pipiens quinquefasciatus (Diptera: Culicidae)
LUO Lan, CUI Feng*
(State Key Laboratory of Integrated Management of Pest Insects and Rodents, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, China)
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Abstract 【Aim】 This study aims to establish and molecularly identify a cell line based on tissues from the newly hatched larvae of the mosquito Culex pipiens quinquefasciatus (Diptera: Culicidae). 【Methods】 The shredded tissues of the newly hatched larvae of the parathion-resistant strain Shengui of C. pipens quinquefasciatus were cultured in M199 complete medium supplemented with 20% heatinactivated fetal bovine serum, and the primary cells were obtained. The primary cells were treated with trypsin and subcultured into a cell line. The tissue origin of the cell line was identified by observing its biological characteristics under a microscope and comparing DNA amplification fingerprinting and sequencing the ribosome DNA internal transcribed spacer. 【Results】 The cell line Cxq-1 was well maintained in M199 complete medium and subcultured continually for 12 months. After being frozen in liquid nitrogen, Cxq-1 was successfully resuscitated for several times. DNA amplification fingerprinting polymerase chain reaction clearly distinguished Cxq-1 from two other insect cell lines. Sequencing of ribosome DNA internal transcribed spacer of Cxq-1 indicated that its rDNA-ITS1 and rDNA-ITS2 sequences has higher than 99% identity with those of C. pipens quinquefasciatus registered in the GenBank, confirming the origin of Cxq-1. 【Conclusion】 The newly established cell line Cxq-1 from newly hatched larvae of C. pipens quinquefasciatus has been confirmed to be a cell line of this mosquito. This cell line provides an important platform for studying the molecular mechanisms of insecticide resistance, mosquito-borne viruses and parasites in the future.
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Key wordsCulex pipiens quinquefasciatus   cell line   DNA amplification fingerprinting   internal transcribed spacer   molecular identification     
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LUO Lan,CUI Feng. Establishment and identification of a new cell line from Culex pipiens quinquefasciatus (Diptera: Culicidae)[J]. ACTA ENTOMOLOGICA SINICA, 2018, 61(1): 79-85.
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Supported by:Beijing Magtech