| 產品名稱 |
Trypanosoma cruzi Chagas |
| 商品貨號 |
B243449 |
| Strain Designations |
CA-I CL72 Lampit Resistant |
| Application |
Vector borne research |
| Biosafety Level |
2
Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country. |
| Isolation |
Lampit (=Nifurtimox) resistant strain derived from CA-I CL72 (=ATCC 50791) |
| Product Format |
frozen |
| Storage Conditions |
Frozen: -70°C or colder for 1 week, vapor phase of liquid nitrogen for long-term storage |
| Axenic/Xenic |
Axenic |
| Type Strain |
no |
| Medium |
ATCC® Medium 1029: LIT medium
|
| Growth Conditions |
Temperature: 20-25°C |
| Cryopreservation |
- Harvest cells from several cultures in very late logarithmic to early stationary phase of growth. Vigorously agitate to suspend the cells.
- Aseptically transfer the cell suspension to 15 ml plastic centrifuge tubes.
- Centrifuge at ~800 x g for 5 min.
- While cells are centrifuging, prepare a 10% solution of DMSO in complete ATCC Medium 1029. Cool on ice.
- Remove the supernatant and pool the cell pellets to the final volume desired with fresh growth medium.
- Combine the cell suspension with an equal volume of 10% DMSO cryoprotectant solution (prepared in step 4) to yield a final concentration of 5% DMSO.
- Dispense in 0.5 ml aliquots to 1.0-2.0 ml Nunc vials (special plastic vials for cryopreservation).
- Place the vials in a controlled rate freezing unit. From room temperature cool at -1°C/min to -40°C. At -40°C, plunge ampules into liquid nitrogen. Alternatively, place the vials in a Nalgene 1°C freezing apparatus. Place the apparatus at -80°C for 1.5 to 2 hours and then plunge ampules into liquid nitrogen. (The cooling rate in this apparatus is approximately -1°C/min.).
- Store ampules in a liquid nitrogen refrigerator until needed.
- To establish a culture from the frozen state, place a frozen ampule in a 35°C water bath just enough to cover the frozen material. Allow the ampule to thaw completely (2-3 min).
- Immediately after thawing, aseptically remove the contents and transfer to a T-25 tissue culture flask containing 10 ml of fresh complete ATCC medium 1029.
- Screw the cap on tightly and incubate at 20-25°C. Observe the culture daily and transfer when numerous trophozoites are observed.
|
| Name of Depositor |
JA Dvorak |
| Special Collection |
NCRR Contract |
| References |
Nozaki T, et al. Cellular and molecular biological analyses of nifurtimox resistance in Trypanosoma cruzi. Am. J. Trop. Med. Hyg. 55: 111-117, 1996. PubMed: 8702014
|
