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ProPak-X.36 [PP-X.36]
ProPak-X.36 [PP-X.36]
規格:
價格:詢價
編號:B165521
品牌:Mingzhoubio

標準菌株
定量菌液
DNA
RNA

規格:
凍干粉
斜面
甘油
平板


產品名稱
ProPak-X.36 [PP-X.36]
商品貨號
B165521
Organism
Homo sapiens, human
Tissue
kidney
培養基
Product Format
frozen
Culture Properties
adherent
Biosafety Level
2 Cells contain adenovirus and CMV viral sequences

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
Leukemia
Age
embryo
Applications
The ProPak packaging cell lines produce either murine leukemia virus (MLV) xenotropic particles (ProPak-X cells; ATCC CRL-12007) or amphotropic particles (ProPak-A cells; ATCC CRL-12006 and ATCC CRL-12479).
They were derived from the human embryonic kidney line, 293 (see ATCC CRL-1573).
The resulting cells were screened for Env expression and clones designated ProPak-X, expressing high levels of Env were screened for ability to produce transducing vector.
ProPak-based producer cells were demonstrated to be free of replication-competent retrovirus (RCR) by stringent testing.
Consistently higher transduction of target cells was achieved with ProPak-derived amphotropic vector than with PA317-packaged amphotropic vector.
Storage Conditions
liquid nitrogen vapor phase
Disclosure
This material is cited in a US or other Patent and may not be used to infringe the claims. Depending on the wishes of the Depositor, ATCC may be required to inform the Patent Depositor of the party to which the material was furnished. This material may not have been produced or characterized by ATCC.
Derivation
They were derived from the human embryonic kidney line, 293 (see ATCC CRL-1573).
The CMV promoter was excised (EcoRI/XhoI, blunt-ended), and replaced with the MoMLV LTR (Asp 718/HindIII, blunt-ended) from plasmid pVH2.
Consistently higher transduction of target cells was achieved with ProPak-derived amphotropic vector than with PA317-packaged amphotropic vector.
Clinical Data
To construct the ProPak-X and the ProPak-A-52 cell lines, the ATG in the splice donor/splice acceptor of pCMV plasmid was mutated to ACG.
Comments
The ProPak packaging cell lines produce either murine leukemia virus (MLV) xenotropic particles (ProPak-X cells; ATCC CRL-12007) or amphotropic particles (ProPak-A cells; ATCC CRL-12006 and ATCC CRL-12479).
They were derived from the human embryonic kidney line, 293 (see ATCC CRL-1573).
To construct the ProPak-X and the ProPak-A-52 cell lines, the ATG in the splice donor/splice acceptor of pCMV plasmid was mutated to ACG.
The CMV promoter was excised (EcoRI/XhoI, blunt-ended), and replaced with the MoMLV LTR (Asp 718/HindIII, blunt-ended) from plasmid pVH2.
The beta-galactosidase gene was replaced by the gag-pol ORF (NotI fragment) to generate pMoMLVgp. pMoMLVgp was co-transfected with pHA58 into 293 cells by calcium phosphate co-precipitation and hygromycin B-resistant cells were selected.
Clones were screened for the level of Gag secretion and one clone secreting high levels of Gag was selected (designated ProGag); this clone yielded high viral titers in transient transfection.
The expression plasmid containing the murine xenotropic env gene, pCI-Ex, was co-transfected with pSV2pac into the ProGag cell line by calcium phosphate precipitation and puromycin-resistant cells were selected.
The resulting cells were screened for Env expression and clones designated ProPak-X, expressing high levels of Env were screened for ability to produce transducing vector. Clone 36 designated ProPak-X.36 was deposited as CRL-12007.
ProPak-based producer cells were demonstrated to be free of replication-competent retrovirus (RCR) by stringent testing.
Consistently higher transduction of target cells was achieved with ProPak-derived amphotropic vector than with PA317-packaged amphotropic vector.
The highest transduction of human hematopoietic progenitor cells was achieved with vector supernatant generated from a coculture of the ProPak-X and ProPak-A cell lines.
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.
  1. Remove and discard culture medium.
  2. Briefly rinse the cell layer with 0.25% (w/v) Trypsin-0.53mM EDTA solution to remove all traces of serum which contains trypsin inhibitor.
  3. 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.
  4. Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting.
  5. Add appropriate aliquots of the cell suspension to new culture vessels.
  6. Incubate cultures at 37°C.

Subcultivation Ratio: 1:4 to 1:10
Medium Renewal: Every 2 to 3 days

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
culture medium 95%; DMSO, 5%
Culture Conditions
Temperature: 37°C
Atmosphere: Air, 95%; Carbon dioxide (CO2), 5%
STR Profile
Amelogenin: X
CSF1PO: 11,12
D13S317: 12,14
D16S539: 9
D5S818: 8,9
D7S820: 11,12
THO1: 7,9.3
TPOX: 11
vWA: 16,19
Name of Depositor
SyStemix, Inc.
Deposited As
human
U.S. Patent Number
References

Rigg RJ, et al. Method for obtaining retroviral packaging cell lines producing high transducing efficiency retroviral supernatant. US Patent 6,017,761 dated Jan 25 2000

Forestell SP, et al. Novel retroviral packaging cell lines: complementary tropisms and improved vector production for efficient gene transfer. Gene Ther. 4: 600-610, 1997. PubMed: 9231077

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