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- 详细信息
- 询价记录
- 文献和实验
- 技术资料
- 品系：
NIH/Swiss
- 生长状态：
贴壁生长
- 物种来源：
小鼠
- 是否是肿瘤细胞：
0
- 相关疾病：
正常
- ATCC Number：
CRL-10686™
- 运输方式：
冻存运输
- 年限：
embryo
- 库存：
大量
- 细胞形态：
成纤维样
| Designations: | PG13 | ||
| Depositors: | Fred Hutchinson Cancer Res. Cntr. | ||
| Biosafety Level: | 2 [CELLS CONTAIN RETROVIRUS ] | ||
| Shipped: | frozen | ||
| Medium & Serum: | See Propagation | ||
| Growth Properties: | adherent | ||
| Organism: | Mus musculus | ||
| Morphology: | fibroblast |
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| Source: | Disease: normal Strain: NIH/Swiss |
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| Permits/Forms: | In addition to the MTA mentioned above, other ATCC and/or regulatory permits may be required for the transfer of this ATCC material. Anyone purchasing ATCC material is ultimately responsible for obtaining the permits. Please click here for information regarding the specific requirements for shipment to your location. | ||
| Applications: | transfection host | ||
| Reverse Transcript: | positive | ||
| Age: | embryo | ||
| Comments: | PG13 is a retrovirus packaging cell line derived from TK- NIH/3T3 cells and based on the Gibbon ape leukemia virus (GaLV). Introduction of retroviral vectors by infection or transfection results in the production of retrovirus virions capable of infecting cells from many species (with the exception of mice). Selection against loss of the DNAs conferring the packaging functions can be performed by growing the cells in medium containing dialyzed fetal bovine serum and 100 nM amethopterin for 5 days. Followed by cultivation in medium containing HAT and untreated fetal bovine serum for an additional 5 days. After selection, the cells should be maintained in medium containing HT for 2 days to avoid toxic effects due to residual amethopterin. A derivative of this line is available (PG13/LN c8, see ATCC CRL-10685 ) that produces a neomycin resistance transducing vector. Although virus production has not been observed, there is a possibility that the cells will produce a virus similar to GaLV (a moderate risk oncogenic virus) and Biosafety Level 2 or higher precautions should be taken when using this cell line. |
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| Propagation: | ATCC 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%. Temperature: 37.0°C |
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| Subculturing: | Protocol: Remove medium, add fresh 0.25% Trypsin, 0.53 mM EDTA solution and allow the flask to sit at room temperature (or 37C) until the cells detach (2 to 3 minutes). Add fresh medium, aspirate and dispense into new flasks. Subcultivation Ratio: A subcultivation ratio of 1:8 is recommended Medium Renewal: Every 2 to 3 days |
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| Preservation: | Freeze medium: Complete growth medium, 95%; DMSO, 5% Storage temperature: liquid nitrogen vapor temperature |
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| Related Products: | Recommended medium (without the additional supplements or serum described under ATCC Medium):ATCC 30-2002 recommended serum:ATCC 30-2020 derivative:ATCC CRL-10685 |
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| References: | 1818: Miller AD, et al. Construction and properties of retrovirus packaging cells based on gibbon ape leukemia virus. J. Virol. 65: 2220-2224, 1991. PubMed: 1850008 4473: Miller AD, Rosman GJ. Improved retroviral vectors for gene transfer and expression. BioTechniques 7: 980-990, 1989. PubMed: 2631796 |
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文献和实验感染 2 h 后换液，然后每天换液继续培养，直到待测组 (未使用病毒感染，只有 PG4 细胞组) 全部长满，此时阳性组 (GALV 病毒感染 2 h 组) 会出现很多空斑，即细胞不能长满。现在问题来了，一直听同事说 PG4 细胞有点奇怪，它不像 PG13 细胞，293T 细胞和 Hela 细胞等在培养皿里是均匀平铺的，而是许多细胞聚集在一起成为一个个细胞群，这样细胞群和细胞群之间本来就有很大的空洞，那怎么来证明这些空洞是因为 GALV 病毒导致的呢？后来同事甚至怀疑这个细胞了，干脆又买了一支细胞，然而新买
Gene Delivery to Cells in Culture Using Retroviruses
Moloney leukemia virus-based vectors can be generated in cells that express the products of three retroviral genes, gag, pol , and env . There are a number of cell lines such as PG13 (1 ) and FLYA13 (2 ), known as packaging cells
、PG13、DA、CFA(第三代包装细胞)，包装细胞可提供逆转录病毒gag、pol和env蛋白才能使带有包装信号及目的基因的病毒载体RNA进行包装，包装细胞只提供gag、pol和env蛋白而不产生具有复制能力的野生型病毒(RCR)，而第一代包装细胞可产生 RCR，安全性较差；第二代包装细胞，临床上已广泛应用，也未发现产生RCR，安全性好；第三代包装细胞更加安全，第三代包装细胞中主要区别是病毒结构基因中env不同。 反转录病毒作为基因转移的载体有如下特点：①反转录病毒感染细胞的效率高，基因
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