Plasmodium falciparum Welch
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ATCC determines the biosafety level of a material based on our risk assessment as guided by the current edition of Biosafety in Microbiological and Biomedical Laboratories (BMBL), U.S. Department of Health and Human Services. It is your responsibility to understand the hazards associated with the material per your organization’s policies and procedures as well as any other applicable regulations as enforced by your local or national agencies.
ATCC highly recommends that appropriate personal protective equipment is always used when handling vials. For cultures that require storage in liquid nitrogen, it is important to note that some vials may leak when submersed in liquid nitrogen and will slowly fill with liquid nitrogen. Upon thawing, the conversion of the liquid nitrogen back to its gas phase may result in the vial exploding or blowing off its cap with dangerous force creating flying debris. Unless necessary, ATCC recommends that these cultures be stored in the vapor phase of liquid nitrogen rather than submersed in liquid nitrogen.
In vitro culture in human erythrocyte
The following directions for recovery from the frozen state must be carefully followed if a culture is to be successfully established.
1. Place the frozen vial in a 37oC water bath until mixture is completely thawed.
2. Aseptically transfer the contents to a 50 ml sterile conical tube.
3. Slowly add 1 volume (0.1 ml) 12% Sodium Chloride solution dropwise via a 1ml syringe to 5 volumes sample (0.5 ml) and agitate continuously.
4. Allow the mixture to stand for 5 mins. at room temperature.
5. Slowly add 5 ml 1.8% Sodium Chloride dropwise via a larger syringe and allow to stand at room temperature for 2 mins.
6. Add 5 ml of 0.9% Sodium Chloride / 0.2% Glucose solution as in step 5.
7. Centrifuge for 5 min. at 1500rpm, remove supernatant.
8. Wash pellet in 20 ml incomplete medium.
9. Centrifuge for 5 min at 1500 rpm, remove supernatant.
10. Resuspend pellet in 8ml complete medium in tissue culture flask and gently aerate culture with gas mixture of 5% CO2, 5% O2 and 90% N2 using a sterile, cotton plugged Pasteur pipet.
11. Smear if required (see below).
1. Centrifuge ring stage culture for 5 min at 1800rpm in 50 ml centrifuge tube.
2. Aspirate supernatant using sterile Pasteur pipet.
3. Resuspend pellet gently in remaining supernatant.
4. Slowly add 5 volumes of glycerolyte medium (see below) to 3 volumes pellet dropwise via a syringe as follows:
A. Add the first volume of glycerolyte and allow the tube to stand for 5 mins. at room temperature.
B. Add the remaining 4 volumes of glycerolyte and gently agitate.
5. Aliquot mixture into Nunc screwtop freezing vials and store at 80oC overnight.
6. Plunge vials into liquid nitrogen (-196oC) the next day and store in liquid nitrogen or liquid nitrogen vapor.
** To formulate glycerolyte medium, combine the following with distilled water to 100 ml: 57.00g glycerol, 1.60g sodium lactate (C3H5NaO3), 30.00mg potassium chloride (KCl), 1.38g sodium dihydrogen phosphate (NaH2PO4). Mix well and adjust pH to 6.8 using concentrated NaOH and/or HCl. Autoclave to sterilize, and store at 4oC.
The product is provided 'AS IS' and the viability of ATCC® products is warranted for 30 days from the date of shipment, provided that the customer has stored and handled the product according to the information included on the product information sheet, website, and Certificate of Analysis. For living cultures, ATCC lists the media formulation and reagents that have been found to be effective for the product. While other unspecified media and reagents may also produce satisfactory results, a change in the ATCC and/or depositor-recommended protocols may affect the recovery, growth, and/or function of the product. If an alternative medium formulation or reagent is used, the ATCC warranty for viability is no longer valid. Except as expressly set forth herein, no other warranties of any kind are provided, express or implied, including, but not limited to, any implied warranties of merchantability, fitness for a particular purpose, manufacture according to cGMP standards, typicality, safety, accuracy, and/or noninfringement.
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Trager W, et al. Clones of the malaria parasite Plasmodium falciparum obtained by microscopic selection: their characterization with regard to knobs, chloroquine sensitivity, and formation of gametocytes. Proc. Natl. Acad. Sci. USA 78: 6527-6530, 1981. PubMed: 7031656