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TIME-GFP
TIME-GFP
規格:
價格:
編號:B208644
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
TIME-GFP
商品貨號
B208644
Organism
Homo sapiens, human
Tissue
foreskin
培養基
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
ATCC CRL-4045 Cell Micrograph
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.
  1. Remove and discard spent medium.
  2. Briefly rinse the cells with Dulbecco's Phosphate Buffered Saline (DPBS, ATCC 30-2200) and discard rinse solution.
  3. 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).
  4. Neutralize trypsin by adding an equal volume of room temperature 2% FBS in DPBS.
  5. Centrifuge cells at 250 x g for 5 min at room temperature.
  6. Remove supernatant. Resuspend pellet in 6.0 to 8.0 mL Complete Growth Medium.
  7. 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
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