BJ-5ta細(xì)胞

細(xì)胞類型： 其他細(xì)胞類型

是否是腫瘤細(xì)胞： 0

物種來源： 人

器官來源： 其他

生長狀態(tài)： 貼壁生長

運(yùn)輸方式： 凍存運(yùn)輸

數(shù)量： 大量

年限： newborn

ATCC Number： CRL-4001?

細(xì)胞形態(tài)： 成纖維樣

Designations: BJ-5ta

Depositors: Geron Corporation

BJ-5ta細(xì)胞Biosafety Level: 1

Shipped: frozen

Medium & Serum: See Propagation

Growth Properties: adherent

Organism: Homo sapiens

Morphology: fibroblast

Source: Organ: skin, foreskin

Cell Type: fibroblast immortalized with hTERT

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.

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Applications: BJ-5ta細(xì)胞TAs 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 firstrecovered 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.

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].

Reverse Transcript: N

Antigen Expression: fibroblast surface protein; Homo sapiens, expressed (fibroblast surface protein (FSP) was assayed by flow cytometry.) (fibroblast surface protein (FSP) was assayed by flow cytometry.)

cytokeratin; Homo sapiens(cytokeratins were assayed by immunocytochemistry using a pan-cytokeratin antibody) (cytokeratins were assayed by immunocytochemistry using a pan-cytokeratin antibody)

DNA Profile (STR): 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

Cytogenetic Analysis: BJ-5ta細(xì)胞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.

Age: newborn

Gender: male

HeLa Markers: N

Comments: 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].

TAs 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 firstrecovered 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.

Propagation: ATCC 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

Atmosphere: air, 95%; BJ-5ta細(xì)胞carbon dioxide (CO2), 5%

Temperature: 37.0°C

Growth Conditions: Subculture when cell concentration reaches between 8 X 10(3) and 1 X 10(4) cells/cm2.

Subculturing: Protocol: Volumes are given for a 75-cm2 flask; proportionally reduce or increase the amount of dissociation medium 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 37C 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 10(3) to 5 x 10(3) viable cells/cm2 is recommended.

Subcultivation Ratio: 1:2 to 1:3 twice weekly

Medium Renewal: every 2 to 3 days

Preservation: Freeze medium: culture medium, 30%; fetal bovine serum, 60%; DMSO, 10%

Storage temperature: liquid nitrogen vapor phase

Doubling Time: 62 hr

Related Products: recommended serum:ATCC 30-2020

Trypsin-EDTA Solution:ATCC 30-2101

Cell culture tested DMSO:ATCC 4-X

Erythrosin B vital stain solution:ATCC 30-2404

Trypan Blue vital stain solution:ATCC 30-2402

plasmid in bacteria:ATCC MBA-141

References: 47354: BJ-5ta細(xì)胞Bodnar AG, et al. Extension of life-span by introduction of telomerase into normal human cells. Science 279: 349-352, 1998. PubMed: 9454332

90421: 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