PA317 ATCC ® CRL-9078™ Mus musculus

Permits	These permits may be required for shipping this product: Customers located in the state of Hawaii will need to contact the Hawaii Department of Agriculture to determine if an Import Permit is required. A copy of the permit or documentation that a permit is not required must be sent to ATCC in advance of shipment.
Organism	Mus musculus, mouse
Product Format	frozen
Morphology	fibroblast
Culture Properties	adherent
Biosafety Level	1
Age	embryo
Strain	NIH/Swiss
Applications	transfection host Packaging line for retroviruses.
Storage Conditions	liquid nitrogen vapor temperature

Derivation	Derived from TK- NIH/3T3 cells by cotransfection with packaging construct DNA, pPAM3, carried in pBR322 and herpes simplex virus thymidine kinase (TK) gene carried in pBR322. The PA317 cell line was derived from TK- NIH/3T3 cells by cotransfection with packaging construct DNA (pPAM3) carried in pBR322 and the herpes simplex virus thymidine kinase (TK) gene carried in pBR322. Virions produced by this line have been used successfully to transfer genes into humans.
Comments	Derived from TK- NIH/3T3 cells by cotransfection with packaging construct DNA, pPAM3, carried in pBR322 and herpes simplex virus thymidine kinase (TK) gene carried in pBR322. The PA317 cell line was derived from TK- NIH/3T3 cells by cotransfection with packaging construct DNA (pPAM3) carried in pBR322 and the herpes simplex virus thymidine kinase (TK) gene carried in pBR322. Introduction of retroviral vectors into these cells, by infection or by transfection, results in production of retrovirus virions with an amphotropic host range that are capable of infecting cells of many mammalian species. Virions produced by this line have been used successfully to transfer genes into humans. The percentage of PA317 cells that are capable of packaging retroviral vectors decreases slowly with continued passaging of the cell line, presumably due to the loss of the transfected DNA used to create the line. Brief selection (about 5 days) in medium containing 0.03 mM hypoxanthine, 0.001 mM amethopterin (methotrexate), and 0.02 mM thymidine will select for cells that retain the packaging function. After selection, the cells should be grown in HT medium (0.03 mM hypoxanthine, 0.02 mM thymidine) for 4 days to dilute any residual amethopterin. Cells shipped from the ATCC have been selected in medium containing 0.03 mM hypoxanthine, 0.001 mM amethopterin (methotrexate), and 0.02 mM thymidine prior to freezing, and should be grown in HT medium for 4 days after receipt. After this, the cells are stable for at least 1 month in the absence of further selection.

Derivation

Derived from TK- NIH/3T3 cells by cotransfection with packaging construct DNA, pPAM3, carried in pBR322 and herpes simplex virus thymidine kinase (TK) gene carried in pBR322.

The PA317 cell line was derived from TK- NIH/3T3 cells by cotransfection with packaging construct DNA (pPAM3) carried in pBR322 and the herpes simplex virus thymidine kinase (TK) gene carried in pBR322.

Virions produced by this line have been used successfully to transfer genes into humans.

Comments

Derived from TK- NIH/3T3 cells by cotransfection with packaging construct DNA, pPAM3, carried in pBR322 and herpes simplex virus thymidine kinase (TK) gene carried in pBR322.

The PA317 cell line was derived from TK- NIH/3T3 cells by cotransfection with packaging construct DNA (pPAM3) carried in pBR322 and the herpes simplex virus thymidine kinase (TK) gene carried in pBR322.

Introduction of retroviral vectors into these cells, by infection or by transfection, results in production of retrovirus virions with an amphotropic host range that are capable of infecting cells of many mammalian species.

Virions produced by this line have been used successfully to transfer genes into humans.

The percentage of PA317 cells that are capable of packaging retroviral vectors decreases slowly with continued passaging of the cell line, presumably due to the loss of the transfected DNA used to create the line.

Brief selection (about 5 days) in medium containing 0.03 mM hypoxanthine, 0.001 mM amethopterin (methotrexate), and 0.02 mM thymidine will select for cells that retain the packaging function.

After selection, the cells should be grown in HT medium (0.03 mM hypoxanthine, 0.02 mM thymidine) for 4 days to dilute any residual amethopterin.

Cells shipped from the ATCC have been selected in medium containing 0.03 mM hypoxanthine, 0.001 mM amethopterin (methotrexate), and 0.02 mM thymidine prior to freezing, and should be grown in HT medium for 4 days after receipt.

After this, the cells are stable for at least 1 month in the absence of further selection.

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	Protocol: Remove medium, and rinse with 0.25% trypsin, 0.03% EDTA solution. Remove the solution and add an additional 1 to 2 ml of trypsin-EDTA solution. Allow the flask to sit at room temperature (or at 37C) until the cells detach. Add fresh culture medium, aspirate and dispense into new culture flasks. Subcultivation Ratio: A subcultivation ratio of 1:10 is recommended Medium Renewal: 2 to 3 times per week
Cryopreservation	Freeze medium: Complete growth medium 95%; DMSO, 5% Storage temperature: liquid nitrogen vapor temperature
Culture Conditions	Temperature: 37.0°C

Name of Depositor	Fred Hutchinson Cancer Res. Cntr.
References	Miller AD, Buttimore C. Redesign of retrovirus packaging cell lines to avoid recombination leading to helper virus production. Mol. Cell. Biol. 6: 2895-2902, 1986. PubMed: 3785217 Miller AD. DNA contructs for retrovirus packaging cell lines. US Patent 4,861,719 dated Aug 29 1989 Rosenberg SA, et al. Gene transfer into humans--immunotherapy of patients with advanced melanoma, using tumor-infiltrating lymphocytes modified by retroviral gene transduction. N. Engl. J. Med. 323: 570-578, 1990. PubMed: 2381442 Giguere V, et al. Identification of a new class of steroid hormone receptors. Nature 331: 91-94, 1988. PubMed: 3267207 Scaglioni PP, et al. Posttranscriptional regulation of hepatitis B virus replication by the precore protein. J. Virol. 71: 345-353, 1997. PubMed: 8985356 Lai CF, et al. Receptors for interleukin (IL)-10 and IL-6-type cytokines use similar signaling mechanisms for inducing transcription through IL-6 response elements. J. Biol. Chem. 271: 13968-13975, 1996. PubMed: 8662928 Roche Transfection Reagents

Name of Depositor

Fred Hutchinson Cancer Res. Cntr.

References

Miller AD, Buttimore C. Redesign of retrovirus packaging cell lines to avoid recombination leading to helper virus production. Mol. Cell. Biol. 6: 2895-2902, 1986. PubMed: 3785217

Miller AD. DNA contructs for retrovirus packaging cell lines. US Patent 4,861,719 dated Aug 29 1989

Rosenberg SA, et al. Gene transfer into humans--immunotherapy of patients with advanced melanoma, using tumor-infiltrating lymphocytes modified by retroviral gene transduction. N. Engl. J. Med. 323: 570-578, 1990. PubMed: 2381442

Giguere V, et al. Identification of a new class of steroid hormone receptors. Nature 331: 91-94, 1988. PubMed: 3267207

Scaglioni PP, et al. Posttranscriptional regulation of hepatitis B virus replication by the precore protein. J. Virol. 71: 345-353, 1997. PubMed: 8985356

Lai CF, et al. Receptors for interleukin (IL)-10 and IL-6-type cytokines use similar signaling mechanisms for inducing transcription through IL-6 response elements. J. Biol. Chem. 271: 13968-13975, 1996. PubMed: 8662928

Roche Transfection Reagents

Permits

These permits may be required for shipping this product:

Customers located in the state of Hawaii will need to contact the Hawaii Department of Agriculture to determine if an Import Permit is required. A copy of the permit or documentation that a permit is not required must be sent to ATCC in advance of shipment.

Basic Documentation

Product Sheet

Certificate of Analysis

MSDS

References

Miller AD, Buttimore C. Redesign of retrovirus packaging cell lines to avoid recombination leading to helper virus production. Mol. Cell. Biol. 6: 2895-2902, 1986. PubMed: 3785217

Miller AD. DNA contructs for retrovirus packaging cell lines. US Patent 4,861,719 dated Aug 29 1989

Rosenberg SA, et al. Gene transfer into humans--immunotherapy of patients with advanced melanoma, using tumor-infiltrating lymphocytes modified by retroviral gene transduction. N. Engl. J. Med. 323: 570-578, 1990. PubMed: 2381442

Giguere V, et al. Identification of a new class of steroid hormone receptors. Nature 331: 91-94, 1988. PubMed: 3267207

Scaglioni PP, et al. Posttranscriptional regulation of hepatitis B virus replication by the precore protein. J. Virol. 71: 345-353, 1997. PubMed: 8985356

Lai CF, et al. Receptors for interleukin (IL)-10 and IL-6-type cytokines use similar signaling mechanisms for inducing transcription through IL-6 response elements. J. Biol. Chem. 271: 13968-13975, 1996. PubMed: 8662928

Roche Transfection Reagents

PA317 (ATCC® CRL-9078™)

Share this product

Your session will expire in

PA317 (ATCC^® CRL-9078^™)