產(chǎn)品名稱 |
BJ-5ta |
商品貨號 |
B218466 |
Organism |
Homo sapiens, human |
Tissue |
Foreskin |
Cell Type |
Fibroblast immortalized with hTERT |
Product Format |
frozen |
Morphology |
Fibroblast-like |
Culture Properties |
Adherent |
Biosafety Level |
1
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 |
Normal |
Age |
Neonatal |
Gender |
Male |
Storage Conditions |
Liquid nitrogen vapor phase |
Karyotype |
This is a diploid human cell line of male origin with a modal chromosome number of 46 that occurred in 90% of the cells counted. The sex chromosomes, X and Y are both karyotypically normal. |
Images |
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Derivation |
The hTERT-immortalized foreskin fibroblast cell line, BJ-5ta, was derived by transfecting the BJ foreskin fibroblast cell line with the pGRN145 hTERT-expressing plasmid (ATCC MBA-141) at population doubling 58. Cells were cultured in medium containing hygromycin B until stable clones were selected [Pubmed: 9454332]. |
Antigen Expression |
Antigen expression: Positive for fibroblast surface protein; Homo sapiens, expressed (fibroblast surface protein (FSP) was assayed by flow cytometry.).
Negative for the pan-cytokeratin epithelial marker; Homo sapiens (cytokeratins were assayed by immunocytochemistry using a pan-cytokeratin antibody). |
Complete Growth Medium |
A 4:1 mixture of Dulbecco's medium and Medium 199 with supplements as follows : 4 parts of Dulbecco's Modified Eagle's Medium containing 4 mM L-glutamine, 4.5 g/L glucose and 1.5 g/L sodium bicarbonate 1 part of Medium 199 Supplemented with: 0.01 mg/ml hygromycin B 10% fetal bovine serum
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Subculturing |
Volumes are given for a 75 cm 2 flask. Recommended use of Corning ® T-75 flasks (catalog #430641). Increase or decrease the amount of dissociation medium needed proportionally for culture vessels of other sizes.
- Remove and discard culture medium.
- Add 3.0 to 5.0 mL of 0.25% trypsin-0.53 mM EDTA solution to the flask and observe cells under an inverted microscope until the cell layer is dispersed (usually within 5 to 15 minutes).
Note: To avoid clumping do not hit or shake 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 10.0 mL of complete growth medium and aspirate cells by gently pipetting.
- Add appropriate aliquots of the cell suspension to new culture vessels. An inoculum of 3 x 103 to 5 x 103 viable cells/cm2 is recommended. Maintain cultures at a cell concentration between 8 x 103 and 1 x 104 cells/cm2.
- Subcultivation ratio: 1:2 to 1:3 twice weekly
- Incubate cultures at 37°C.
Subcultivation Ratio: 1:2 to 1:3 twice weekly
Medium Renewal: every 2 to 3 days
Note: Subculture when cell concentration reaches between 8 X 103 and 1 X 104 cells/cm2.
Note: For more information on enzymatic dissociation and subculturing of cell lines consult Chapter 13 in Culture Of Animal Cells: A Manual of Basic Technique by R. Ian Freshney, 5th edition, published by Wiley-Liss, N.Y., 2005. |
Cryopreservation |
Freeze medium: culture medium, 30%; fetal bovine serum, 60%; DMSO, 10% Storage temperature: liquid nitrogen vapor phase |
Culture Conditions |
Atmosphere: air, 95%; carbon dioxide (CO2), 5%
Temperature: 37°C |
STR Profile |
Amelogenin: X,Y CSF1PO: 10,12 D13S317: 8,9 D16S539: 9,13 D5S818: 12 D7S820: 11,12 THO1: 7,8 TPOX: 10,11 vWA: 16,18 |
Population Doubling Level (PDL) |
As part of our quality control, we have tested this cell line for its ability to grow for a minimum of 15 population doublings after recovery from cryopreservation. We have also compared its karyotype, telomerase expression level, growth rate, morphology and tissue-specific markers when first recovered from cryopreservation with that of cells at 10+ population doublings to ensure that there is no change in these parameters and that the cells are capable of extended proliferation. |
Name of Depositor |
Geron Corporation |
Deposited As |
Homo sapiens |
References |
Bodnar AG, et al. Extension of life-span by introduction of telomerase into normal human cells. Science 279: 349-352, 1998. PubMed: 9454332
Jiang XR, et al. Telomerase expression in human somatic cells does not induce changes associated with a transformed phenotype. Nat. Genet. 21: 111-114, 1999. PubMed: 9916802
Aiastui A, et al. Salmonella enterica serovar typhimurium invades fibroblasts by multiple routes differing from the entry into epithelial cells. Infect Immun 78(6):2700-13, 2010. PubMed: 20368348
Janeckova L, et al. HIC1?Tumor?Suppressor?Loss?Potentiates?TLR2/NF-κB?Signaling?and?Promotes?Tissue?Damage-Associated?Tumorigenesis. Mol Cancer Res?13(7):1139-48, 2015. PubMed: 25934696
Amadori S, et al. Effect of sterilization and crosslinking on gelatin films. J Mater Sci Mater Med 26(2):69, 2015. PubMed: 25631265
La Gatta A, et al. Hyaluronan dermal fillers via crosslinking with 1,4-butandiol diglycidyl ether: Exploitation of heterogeneous reaction conditions. J Biomed Mater Res B Appl Biomater 104(1):9-18, 2016. PubMed: 25611588
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