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微信小章| 产品名称： | Ker-CT |
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| 商品货号： | TS140002 |
| Organism： | Homo sapiens, human |
| Tissue： | foreskin |
| Cell Type： | keratinocyte |
| Product Format： | frozen |
| Morphology： | epithelial |
| Culture Properties： | adherent |
| Biosafety Level： | 2 xa0Cells contain SV40 viral DNA 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： | normal |
| Age： | neonatal |
| Gender： | male |
| Applications： | Overexpression of CDK4 and hTERT in neonatal foreskin keratinocyte induced a dramatic upregulation of p16INK4a and milder upregualtion of p53 and p21WAF1, which became unresponsive to UV irradiation. xa0Despite the high levels of these checkpoint factors, Ker-CT cells divide in an apparently normal regulated fashion, are able to respond to changes in calcium levels, retain the stem cell phenotype, and fully differentiate and stratify in organotypic culturexa0Ref |
| Karyotype： | Cytogenetic analysis was performed on G-banded metaphase cells from the human cell line Ker-CT and two abnormal male clones were detected. Clone 1 demonstrated trisomy 5 and trisomy 20. Clone 2 demonstrated trisomy 5 and four copies of chromosome 20. |
| Images： | 
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| Derivation： | The Ker-CT cell line was immortalized by human telomerase and CDK4 using retroviral pBABE-puro hTERT and pSRalphaMSU expressing mouse CDK4. |
| Clinical Data： | male neonatal |
| Antigen Expression： | Positive for p63 (TP63) and Cytokeratin 5 (KRT5) |
| Comments： | The Ker-CT cell line is positive for telomerase, failed to senesce, and was proliferating after more than 250 population doublings Ref  Ramirez R, et al. Bypass of telomere-dependent replicative senescence (M1) upon overexpression of Cdk4 in normal human epithelial cells. Oncogene 22(3): 433-44, 2003. PubMed: 12545164xa0xa0 |
| Complete Growth Medium： | KGM-Gold™ BulletKit™(Lonza 00192060)
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| Subculturing： | Volumes used in this protocol are for 75 cm2 flasks; proportionally reduce or increase amount of dissociation solutions for culture vessels of other sizes.
Medium Renewal: Every 2-3 days.xa0 As the cells become more confluent, increase the volume of media as follows: under 25% confluence feed cells 5 mL per 25 cm2, 25-45% confluence then feed cells 7.5 mL per 25 cm2, over 45% confluence then feed cells 10 mL per 25 cm2. |
| Cryopreservation： | 90% FBS, 10% DMSO |
| Culture Conditions： | Atmosphere: air, 95%; carbon dioxide (CO2), 5% Temperature: 37°C |
| STR Profile： | Amelogenin: X,Y CSF1PO: 7,11 D13S317: 11,13 D16S539: 9,11 D5S818: 12 D7S820: 10,11 TH01: 6,9 TPOX: 8 vWA: 15,16 |
| Name of Depositor： | J Shay |
| References： | Ramirez R, et al. Bypass of telomere-dependent replicative senescence (M1) upon overexpression of Cdk4 in normal human epithelial cells. Oncogene 22(3): 433-44, 2003. PubMed: 12545164 Vaughan M, et al. H-ras expression in immortalized keratinocytes produces an invasive epithelium in cultured skin equivalents. PLoS One 4(11): e7908, 2009. PubMed: 19936293 |