MediaDive DSMZ
Content
Media Solutions Ingredients Strains Steps Gas
Search
Medium finder Solution finder Taxonomy search Isolation sources
Tools
Medium builder NEW Unit converter Compare media Compare solutions Prediction SOON
DSMZ
Back to all solutions

Main sol. 1555

Download this solution as

CSV JSON PDF

Internal Solution-ID: S3212

1 Cultivation of IRE11 cells for infection with Rickettsia monacensis :
2 Alterations were made to the following reagents:
3 L15C (Modified Leibovitzs' L15) used to culture host cells (IRE11). L15C medium is a modification of L15B medium, previously described in detail (Munderloh UG, Kurtti TJ. 1989. Formulation of medium for tick cell culture. Exp. Appl. Acarol. 7:219-29).
Compound Amount Unit Conc. [g/L] Conc. [mM]
L-Aspartic acid 0.4485 g/L 0.449 3.37
L-Glutamine 0.5000 g/L 0.5 3.421
L-Proline 0.4500 g/L 0.45 3.909
L-Glutamic acid 0.2500 g/L 0.25 1.699
alpha-ketoglutaric acid 0.4485 g/L 0.449 3.07
D-Glucose 18.0180 g/L 18.018 100.011
NaOH 5.0000 mM 0.2 5
4 Prior to use in supplemented medium, L15C was diluted by the addition of 33 % water by volume.
5 Supplemented L15C medium is used with
Fetal bovine serum 10.0000 % 100 -
Tryptose-phosphate broth 5.0000 % 50 -
lipoprotein concentrate
(LPC; MPBiomedical) 		0.1000 % 1 -
HEPES 25.0000 mM 5.958 25
NaHCO3 0.2500 % 2.5 29.759
6 The pH is adjusted to pH 7.5 using 1 M NaOH.
7 This medium and the cultivation of tick cells is described in the "Basic Tick Cell Culture Methods" provided as pdf-file (see strain entry for DSM 29017: cultivation conditions) by Tim Kurtti & Uli Munderloh, University of Minnesota.
8 Infection with Rickettsia monacensis or R. buchneri : One 25 cm2 flask of well-grown cells is infected with 1 ml of the Rickettsia strain (quickly thawed from -80°C ) and centrifuged for 1 h onto the cell layer at 1600 rpm at 20°C . Incubate at 30°C with closed lid (without CO2) and look for bacteria and destroyed cells after 4-6 days .
1555 Medium for tick cells (Rickettsia)