| 產品名稱 |
Fugu fry |
| 商品貨號 |
B186291 |
| Organism |
Fugu niphobles, kusafugu |
| Tissue |
fry, whole |
| Product Format |
frozen |
| Morphology |
fibroblast |
| Culture Properties |
adherent |
| Biosafety Level |
1
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. |
| Disease |
normal |
| Age |
1 to 6 days posthatching juvenile |
| Applications |
The Fugu fry cell line was established in 1996 from normal juvenile whole fry. The cell line may be used for in vitro vertebrate genome studies, and in marine fish and aquaculture research. |
| Storage Conditions |
liquid nitrogen vapor phase |
| Karyotype |
hyperploid |
| Derivation |
The Fugu fry cell line was established in 1996 from normal juvenile whole fry. |
| Comments |
The Fugu fry cell line was established in 1996 from normal juvenile whole fry. Fugu cells maintain an unusually compact genome size with small introns. They are telomerase positive. The cell line may be used for in vitro vertebrate genome studies, and in marine fish and aquaculture research. |
| Complete Growth Medium |
These cells are grown in
- 67.5% DMEM HG without sodium bicarbonate (Invitrogen Cat no. 12100)
- 25% L-15 (ATCC Cat no.30-2008)
- 7.5% Ham?s F12 without sodium bicarbonate (Invitrogen Cat no. 21700)
Supplemented with:
- 0.5 g/L sodium bicarbonate
- 15 mM HEPES
- 0.01 mg/ml bovine insulin
- 0.05 mM 2-mercaptoethanol
- 1.0 mM L-glutamine
- 0.05 mM non-essential amino acids
- 10 ng/ml basic human recombinant FGF
- 50 ng/ml mouse EGF (Do not filter)
- 5 to 7.5% heat-inactivated fetal bovine serum
|
| Subculturing |
Protocol: - Remove and discard culture medium.
-
Briefly rinse the cell layer with 0.25% (w/v) Trypsin- 0.53 mM EDTA solution to remove all traces of serum that contains trypsin inhibitor.
-
Add 2.0 to 3.0 ml of Trypsin-EDTA solution to flask and observe cells under an inverted microscope until cell layer is dispersed (usually within 5 to 15 minutes).
Note: To avoid clumping do not agitate the cells by hitting or shaking the flask while waiting for the cells to detach. Cells that are difficult to detach may be placed at 21 to 23°C to facilitate dispersal. -
Add 6.0 to 8.0 ml of complete growth medium and aspirate cells by gently pipetting.
-
To remove trypsin-EDTA solution, transfer cell suspension to centrifuge tube and spin at approximately 125 xg for 5 to 10 minutes.
-
Discard supernatant and resuspend cells in fresh growth medium. Add appropriate aliquots of cell suspension to new culture vessels.
- Incubate cultures at 21 to 23°C without CO2.
Subcultivation Ratio: A subcultivation ratio of 1:3 to 1:4 is recommended; cell density must be high. Medium Renewal: Every 2 to 3 days |
| Cryopreservation |
Freeze medium: Complete growth medium 95%; DMSO, 5% Storage temperature: liquid nitrogen vapor phase |
| Culture Conditions |
Temperature: 22.0°C; (Max. 23.0 °C, Min. 21.0°C) |
| Population Doubling Time |
2.5 days |
| Name of Depositor |
DW Barnes |
| Deposited As |
Fugu niphobles |
| Year of Origin |
1996 |
| References |
Bradford CS, et al. Characterization of cell cultures derived from Fugu, the Japanese pufferfish. Mol. Mar. Biol. Biotechnol. 6: 279-288, 1997. PubMed: 9418286
|
