Plasmodium berghei Vincke and Lips (ATCC® 50175)

Organism: Plasmodium berghei Vincke and Lips  /  Depositor: NE Alger

Strain Designations NK65A
Application
Vector borne research
Biosafety Level 1
Isolation
derived from M. Yoeli strain NK65 by mosquito passage, Univ. Illinois, Urbana, pre-1978
Product Format frozen
Type Strain no
Growth Conditions
Duration: in vivo, mouse
Protocol: ATCCNO: 50173 SPEC: This strain is distributed as a frozen stabilate. See general instructions for thawing and storage of frozen material before proceeding. Upon arrival remove the frozen ampoule from the dry ice and transfer directly to a 35C water bath. When completely thawed, aseptically remove the material with a syringe and inject the entire thawed contents of the ampule intraperitoneally into a 6- to 9-week-old mouse. The strain should be monitored, especially after infecting with thawed material. To infect additional mice, draw a drop of tail blood into a 1.0 ml syringe containing 0.5 ml of sterile buffered anticoagulant, mix, and inoculate 0.1-0.2 ml of the suspension intraperitoneally/rat.
Subcultivation
Protocol: ATCCNO: 50173 SPEC: This strain is distributed as a frozen stabilate. See general instructions for thawing and storage of frozen material before proceeding. Upon arrival remove the frozen ampoule from the dry ice and transfer directly to a 35C water bath. When completely thawed, aseptically remove the material with a syringe and inject the entire thawed contents of the ampule intraperitoneally into a 6- to 9-week-old mouse. The strain should be monitored, especially after infecting with thawed material. To infect additional mice, draw a drop of tail blood into a 1.0 ml syringe containing 0.5 ml of sterile buffered anticoagulant, mix, and inoculate 0.1-0.2 ml of the suspension intraperitoneally/rat.
Cryopreservation

CRYOPRESERVATION: 

Only young cells (rings) can be frozen in glycerolyte medium* because their membranes are more robust.

1.   To harvest parasites, inject host with ketamine (0.1–0.2 ml).

2.   Open chest cavity to expose heart and exsanguinate via cardiac puncture using Yaeger's anticoagulant** (see below), 1 volume anticoagulant to 4 volumes blood.                                                                                                                                                             Glass distilled HGla                                                                                                                                                                             

3.   Centrifuge blood for 5 mins. at 1800 rpm in 50 ml centrifuge tube.

4.   Aspirate supernatant using sterile Pasteur pipet.

5.   Resuspend pellet gently in remaining supernatant.

6.   Slowly add 5 volumes of glycerolyte medium 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.

7.   Aliquot mixture into Nunc screw-capped freezing vials and place in a Nalgene 1°C cooling apparatus. Place the apparatus at -80°C overnight and then plunge ampules into liquid nitrogen.  (The cooling rate in this apparatus is approximately -1°C/min.).

8.   Plunge vials into liquid nitrogen (-196oC) the next day and store in liquid nitrogen or liquid nitrogen vapor.

       

Name of Depositor NE Alger
Year of Origin 1978
References

Branton MBStudies of clonal populations of NK65 Plasmodium berghei Ph.D. thesis, Univ. Illinois, 1978