1514
1514 .
Grace's Insect Medium
This medium was extracted automatically and has not been curated by experts, yet.
Culture medium recipe
Vol.
×
Main sol. 1514 #
ml
Equipment needed: Filter
| 1 Medium for maintenance of SF-9 cells (ACC 125) for cultivation of Rhabdochlamydiae. | |||||
| Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] | |
|---|---|---|---|---|---|
|
Grace's Insect Medium
(Sigma Aldrich, G8142) |
900 | ml | - | - | |
|
Fetal bovine serum
(heat-inactivated, 56°C, 30 min) |
100 | ml | - | - | |
| 2 Filter-sterilize | |||||
| 3 Maintenance conditions of SF-9 cells (see also DSMZ catalogue for ACC 125): | |||||
| 4 SF-9 cells are cultivated at 25 - 29°C without the addition of CO2 in flasks (25 cm2) with tightly closed lids. When a confluent layer has formed, infection can be carried out. | |||||
| 5 Infection with Rhabdochlamydiae: | |||||
| 6 Exchange medium and add 500 – 1000 µl of EB stock solution (thawed quickly to 37°C). Centrifuge for 1 h onto the cell layer at 1600 rpm at 20°C. | |||||
| 7 Maintenance/Passage of infected insect cell cultures: | |||||
| 8 Infected cultures can be maintained e.g. in 25 cm2 flasks (e.g. Nunc flasks, VWR, 734- 2081) at 27°C in an incubator (no special requirements concerning CO2 or light). If the percentage of infected cells is still low (which might occur initially after inoculation of cultures with bacteria from a frozen stock) infected cultures may be maintained by weekly passage (1:5). Infection of yet uninfected cells by added bacteria or bacteria released from neighboring infected cells is facilitated by centrifugation (130 x g, 15 min) of the culture flask. In well-infected cultures host cell lysis will be visible (the bacteria cause host cell lysis within approximately 5-7 days after invasion of host cells). To maintain such cultures, bacteria have to be transferred by adding culture supernatant (approximately 0.5-1 ml) to fresh culture flasks containing uninfected insect cells (approximately semi-confluent culture) each 7-10 days. | |||||
Solution occurs in 2
other media (first defined in Medium 1500).
| 1 Bacteria released from host cells can be stored in | |||||
| Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] | |
|---|---|---|---|---|---|
| Sucrose-Phosphate-Glutamate buffer | 1000 | ml | - | - | |
| 2 at -80°C for several months. | |||||
| 3 Infected cell cultures (supplemented with 10% DMSO) are best stored in liquid nitrogen. | |||||
Solution occurs in 2
other media (first defined in Medium 1500).
| Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] |
|---|---|---|---|---|
| Sucrose | 75.00 | g | 75 | 219.106 |
| KH2PO4 | 0.52 | g | 0.52 | 3.821 |
| Na2HPO4 x 2 H2O | 1.53 | g | 1.53 | 10.778 |
| Glutamic acid | 0.75 | g | 0.75 | 5.098 |
| Distilled water | 1000.00 | ml | - | - |
Metadata
| Last modified: | 24.02.22 |
| Source: | DSMZ |
| Taxonomic range: | Bacteria |
| Medium type: | Complex medium |
| Final pH: | n.d. |
| Equipment needed: | Filter |
| Edit in Medium builder | |
Associated strains
| All strains for this medium: | 1 |
Molecular composition
| Compound | Conc. [g/L] | Conc. [mM] |
|---|---|---|
| Fetal bovine serum | - | - |
| Grace's Insect Medium | - | - |
