產(chǎn)品名稱 |
TIME-GFP |
商品貨號 |
B208644 |
Organism |
Homo sapiens, human |
Tissue |
foreskin |
Cell Type |
microvascular endothelial cell |
Product Format |
frozen |
Morphology |
endothelial |
Culture Properties |
adherent |
Biosafety Level |
2 [Cells 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 |
Both TIME-GFP (ATCC CRL-4045) and its parental cell line TIME (ATCC CRL-4025) exhibit normal diploid karyotype, extended lifespan in culture and endothelial characteristics that make them an ideal model for investigating many aspects of endothelial cell biology. TIME-GFP (ATCC CRL-4045) retains angiogenic potential and stably expresses green fluorescence protein (GFP) for at least 12 passages. The GFP-expressing cells migrate and coalesce into networks of vessel-like structure within 8 hours after being plated onto basement membrane gel (CellMatrix, ATCC ACS-3035). The stable expression of GFP in these cells enables detection and analysis of the fragile endothelial structures to occur without post-assay fixation and/or staining. |
Shipping Information |
FZ |
Storage Conditions |
liquid nitrogen vapor phase |
Karyotype |
This is a diploid cell line of male origin with a modal chromosome number of 46 and a low rate of polyploidy. The line shows some karyotypic instability at later passages. |
Images |
 |
Derivation |
TIME-GFP (ATCC CRL-4045) is derived by transfecting TIME (ATCC CRL-4025) cells with linearized pWE2-EmGFP plasmid, which constitutively expresses the Green Fluorescence Protein (GFP) driven by a CMV promoter. A stable GFP-expressing clone is selected to establish the TIME-GFP cell line. |
Clinical Data |
male
neonatal |
Antigen Expression |
Positive for CD31 and capable of uptaking Low Density Lipoprotein (LDL). Express Green Fluorescence Protein (GFP) constitutively |
Complete Growth Medium |
The base medium for this cell line is Vascular Cell Basal Medium (ATCC PCS-100-030), supplemented with Microvascular Endothelial Cell Growth Kit-BBE (ATCC PCS-110-040) OR Microvascular Endothelial Cell Growth Kit-VEGF (ATCC PCS-110-041). Add Blasticidin (Life Technologies Cat. No. A11139-03) at a final concentration of 12.5 ug/mL and G418 (Life Technologies Cat. No. 11811-023) at a final concentration of 200 ug/mL. Note: Do not filter complete medium.
|
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.
Subculture when the culture is about 90% confluent.
- Remove and discard spent medium.
- Briefly rinse the cells with Dulbecco's Phosphate Buffered Saline (DPBS, ATCC 30-2200) and discard rinse solution.
- Add 2.0 to 3.0 mL room temperature Trypsin-EDTA for Primary Cells (ATCC PCS-999-003) to the flask. Incubate at 37°C for 5 min (until cells have detached).
- Neutralize trypsin by adding an equal volume of room temperature 2% FBS in DPBS.
- Centrifuge cells at 250 x g for 5 min at room temperature.
- Remove supernatant. Resuspend pellet in 6.0 to 8.0 mL Complete Growth Medium.
- Count cells, and seed 5.0 x 103 to 8.0 x 103 viable cells/cm2 to new culture vessels.
Medium Renewal: Every 2-3 days. |
Cryopreservation |
90% FBS, 10% DMSO |
Culture Conditions |
Atmosphere: air, 95%; carbon dioxide (CO2), 5%
Temperature: 37°C |
STR Profile |
D5S818: 11
D13S317: 9, 11
D7S820: 8, 9 ;
D16S539: 9, 12
vWA: 16, 18
Amelogenin: X, Y
TPOX: 8
CSF1PO: 11, 12
TH01: 6, 7 |
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 |
ATCC |