產(chǎn)品名稱 |
DSDh |
商品貨號 |
B164372 |
Organism |
Canis familiaris, dog |
Tissue |
bone |
Product Format |
frozen |
Morphology |
fibroblast |
Culture Properties |
adherent |
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. |
Disease |
osteosarcoma |
Age |
11 years |
Gender |
female |
Strain |
poodle |
Applications |
DSDh cells were isolated after selection with methotrexate and screening for helper activity. The line is used as a helper cell for propagating replication incompetent vectors derived from spleen necrosis virus (SNV). After serial passaging for 2 months, the cells appears to decrease in helper activity (as evidence by drops in viral titers produced), thus it is best to prepare generous frozen stocks of the cells upon receipt. |
Derivation |
DSDh cells were isolated after selection with methotrexate and screening for helper activity. The line is used as a helper cell for propagating replication incompetent vectors derived from spleen necrosis virus (SNV). After serial passaging for 2 months, the cells appears to decrease in helper activity (as evidence by drops in viral titers produced), thus it is best to prepare generous frozen stocks of the cells upon receipt. |
Clinical Data |
female |
Comments |
DSDh is a retrovirus packaging cell line derive from the D-17 canine osteogenic sarcoma cell line (ATCC CCL-183) by Howard Temin. D-17 cells were transfected with plasmids pBR1 (gag - pol genes from spleen necrosis virus), pPR102 (env gene from spleen necrosis virus) and pFR400 (dihydrofolate reductase gene). DSDh cells were isolated after selection with methotrexate and screening for helper activity. The line is used as a helper cell for propagating replication incompetent vectors derived from spleen necrosis virus (SNV). After serial passaging for 2 months, the cells appears to decrease in helper activity (as evidence by drops in viral titers produced), thus it is best to prepare generous frozen stocks of the cells upon receipt. |
Complete Growth Medium |
Minimum essential medium (Eagle) with Earle's BSS containing 500 nM methotrexate, 92%; fetal bovine serum, 8%
|
Subculturing |
Medium Renewal: Twice per week Remove spent medium, add fresh 0.25% trypsin, 0.03% EDTA solution, rinse and remove trypsin. Add fresh trypsin solution (1 to 2 ml) and let the culture sit at room temperature (or at 37C) until the cells detach. Add fresh medium, aspirate and dispense into new flasks. Subculture at or prior to becoming confluent. Inoculate new flasks with 1 to 2 X 10 exp5 cells per sq cm. |
Cryopreservation |
Culture medium, 95%; DMSO, 5% |
Name of Depositor |
H Temin, DW Burns |
Deposited As |
Canis familiaris |
References |
Hu WS, Temin HM. Genetic consequences of packaging two RNA genomes in one retroviral particle: pseudodiploidy and high rate of genetic recombination. Proc. Natl. Acad. Sci. USA 87: 1556-1560, 1990. PubMed: 2304918
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