| 產品名稱 |
GC-2spd(ts) |
| 商品貨號 |
B164497 |
| Organism |
Mus musculus, mouse |
| Cell Type |
spermatocyte; SV40 large T antigen transfected |
| Product Format |
frozen |
| Morphology |
epithelial |
| Culture Properties |
adherent |
| Biosafety Level |
2 [CELLS CONTAIN PAPOVAVIRUS]
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. |
| Age |
6 weeks |
| Gender |
male |
| Strain |
BALB/c |
| Applications |
This cell line may be useful for the study of spermatogenesis. |
| Storage Conditions |
liquid nitrogen vapor phase |
| Derivation |
GC-2spd(ts) was established by stable cotransfection of freshly isolated spermatocytes with the SV40 large T antigen gene (pSV3neo, see ATCC 37150) and a temperature sensitive mutant of the p53 tumor suppressor gene (LTRp53cG9).
Cells were selected with G-418, cultivated for 6 months and single cell cloned three times by limiting dilution.
No clonal proliferation was observed in soft agar cultures, indicating that these cells were immortalized but not transformed.
|
| Clinical Data |
male |
| Comments |
The cells have lost their differentiation potential, and are currently arrested at a premeiotic stage. |
| Complete Growth Medium |
The base medium for this cell line is ATCC-formulated Dulbecco's Modified Eagle's Medium, Catalog No. 30-2002. To make the complete growth medium, add the following components to the base medium: fetal bovine serum to a final concentration of 10%.
|
| Subculturing |
Volumes used in this protocol are for 75 cm2 flask; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes.
- 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 37°C to facilitate dispersal.
- Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting.
- Add appropriate aliquots of the cell suspension to new culture vessels.
- Incubate cultures at 37°C
Subculture Ratio: 1:5 to 1:7
Medium Renewal: 2 to 3 times a week.
Note: For more information on enzymatic dissociation and subculturing of cell lines consult Chapter 10 in Culture of Animal Cells, a manual of Basic Technique by R. Ian Freshney, 3rd edition, published by Alan R. Liss, N.Y., 1994.
|
| Cryopreservation |
Freeze medium: Complete growth medium, 95%; DMSO, 5% Storage temperature: liquid nitrogen vapor temperature |
| Culture Conditions |
Temperature: 37°C |
| Name of Depositor |
MC Hofmann, J Millan |
| Deposited As |
Mus musculus |
| References |
Hofmann MC, et al. A haploid and a diploid cell coexist in an in vitro immortalized spermatogenic cell line. Dev. Genet. 16: 119-127, 1995. PubMed: 7736662
Hofmann MC, et al. Immortalized germ cells undergo meiosis in vitro. Proc. Natl. Acad. Sci. USA 91: 5533-5537, 1994. PubMed: 8202522
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