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Product Name:
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Sua 4.0 , Anopheles gambiae Cell Line
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Manufacturer:
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BEI Resources
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Material Provided:
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Each vial of MRA-921 contains approximately 0.5 mL of Sua 4.0 cells in Schneider’s Insect medium (protect from light) supplemented with 10% fetal bovine serum (FBS) and 10% dimethylsulfoxide (DMSO). Sufficient cells are provided to initiate at least one new culture. The cell count, expressed as cells /vial, is shown on individual certificates of analysis for each product lot.
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Packing/Storage:
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This product was packaged aseptically in cryovials. It should be stored at -100°C or colder, preferably in the vapor phase of a liquid nitrogen freezer. Storage at -70°C will result in loss of viability. To ensure the highest level of viability, the vial should be thawed and the culture initiated as soon as possible upon receipt. Any warming of the product during shipping and transfer must be avoided, as this will adversely affect the viability of the product after thawing. For transfer between freezers and shipping, the cells may be placed on dry ice for brief periods, although use of a portable liquid nitrogen carrier is preferred. Please read the following recommendations prior to reconstituting this material.
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Safety Precautions:
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When handling frozen vials, it is highly recommended that protective gloves, lab coat and full-face mask be worn. Even brief exposure to the ultra-cold temperature can cause tissue damage from frostbite. Also, some vials may slowly fill with liquid nitrogen if they have been immersed during cryogenic storage. When thawing, the liquid nitrogen may rapidly expand as it changes to gas, breaking the vial or cap with explosive force, sending debris flying with enough velocity to cause injury. Store and use in areas with adequate ventilation.
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Growth Conditions:
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Growth Conditions:
Prior to thawing the cells, prepare media:
NOTE: Schneider’s Insect growth medium is light-sensitive. Please handle the media and the cell line culture in the dark. Failure to do so may result in the death of the cells.
Culture Medium:
• Schneider’s Insect medium (protect from light)
• 10% FBS (qualified for insect cell culture or heat-inactivated) • 100 U/mL Penicillin • 100 µg/mL Streptomycin
Freezing Medium:
• Schneider’s Insect medium (protect from light)
• 10% FBS (qualified for insect cell culture or heat-inactivated) • 10% DMSO
Thaw 1 vial in a 25°C water bath and transfer the contents into a 25 cm2 vented cell culture flask with 9 mL of culture medium. Keep the flask tightly capped in a 27°C to 28°C incubator (no CO2 required). Change the media at 12 to 16 hours post seeding. Feed the cells at least every 48 hours, harvest at 80% to 90% confluency and reseed at a split ratio of 1:2 to 1:4.
Subcultivation Procedure: When cells near confluency, detach cells by vigorous shaking, mechanical disruption or gentle cell scraping. Collect and gently aspirate several times with a pipette to disrupt clumped cells prior to cell counting as required and passage to new flasks.
Note: Trypsin or trypsin-like enzyme substitute may be used to fully disperse adherent cells but is not recommended on a continuous basis for MRA-921.
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Disclaimers:
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You are authorized to use this product for research use only. It is not intended for human use. Use of this product is subject to the terms and conditions of the BEI Resources Material Transfer Agreement (MTA). The MTA is available on our Web site at www.beiresources.org. While BEI Resources uses reasonable efforts to include accurate and up-to-date information on this product sheet, neither ATCC® nor the U.S. Government makes any warranties or representations as to its accuracy. Citations from scientific literature and patents are provided for informational purposes only. Neither ATCC® nor the U.S. Government warrants that such information has been confirmed to be accurate. This product is sent with the condition that you are responsible for its safe storage, handling, use and disposal. ATCC® and the U.S. Government are not liable for any damages or injuries arising from receipt and/ or use of this product. While reasonable effort is made to ensure authenticity and reliability of materials on deposit, the U.S. Government, ATCC®, their suppliers and contributors to BEI Resources are not liable for damages arising from the misidentification or misrepresentation of products. |
References:
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1. Christophides, G. K., Personal Communication.2. Müller, H.-M., et al. “A Hemocyte-Like Cell Line Established from the Malaria Vector Anopheles gambiae Expresses Six Prophenoloxidase Genes.” J. Biol. Chem. 274 (1999): 11727-11735. PubMed: 10206988.
3. Dimopoulos, G., et al. “Genome Expression Analysis of Anopheles gambiae: Responses to Injury, Bacterial Challenge, and Malaria Infection.” Proc. Natl. Acad. Sci. USA 99 (2002): 8814-8819. PubMed: 12077297.
4. Dimopoulos, G., et al. “Anopheles gambiae Pilot Gene Discovery Project: Identification of Mosquito Innate Immunity Genes from Expressed Sequence Tags Generated from Immune-Competent Cell Lines.” Proc. Natl. Acad. Sci. USA 97 (2000): 6619-6624. PubMed: 10841561.
5. Catteruccia, F., et al. “Toward Anopheles Transformation: Minos Element Activity in Anopheline Cells and Embryos.” Proc. Natl. Acad. Sci. USA 97 (2000): 2157-2162. PubMed: 10681436.
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Citation:
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Acknowledgment for publications should read "The following reagent was obtained through BEI Resources, NIAID, NIH: Sua 4.0, Anopheles gambiae Cell Line, MRA-921, contributed by George K. Christophides.”
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Biosafety Level:
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1
Appropriate safety procedures should always be used with this material. Laboratory safety is discussed in the following publication: U.S. Department of Health and Human Services, Public Health Service, Centers for Disease Control and Prevention, and National Institutes of Health. Biosafety in Microbiological and Biomedical Laboratories (BMBL). Current Edition. Washington, DC: U.S. Government Printing Office.
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