CHON-001細(xì)胞

運(yùn)輸方式： 凍存運(yùn)輸

器官來(lái)源： 骨

年限： 18 weeks fetus

ATCC Number： CRL-2846?

相關(guān)**： 正常

細(xì)胞形態(tài)： 成纖維樣

生長(zhǎng)狀態(tài)： 貼壁生長(zhǎng)

數(shù)量： 大量

組織來(lái)源： cartilage

細(xì)胞類型： 其他細(xì)胞類型

是否是腫瘤細(xì)胞： 0

物種來(lái)源： 人

Designations: CHON-001

CHON-001細(xì)胞Depositors: ATCC

Biosafety Level: 2 [cells containing SV40 viral DNA sequences ]

Shipped: frozen

Medium & Serum: See Propagation

Growth Properties: adherent

Organism: Homo sapiens

Morphology: fibroblast-like

Source: Organ: bone

Tissue: cartilage

Disease: normal

Cell Type: fibroblast immortalized with hTERT

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Isolation: Isolation date: December 20, 2001

Applications: CHON-001細(xì)胞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.

The primary cells were infected by the defective retrovirus containing hTERT gene under G418 selection.

The chondrocyte cell line, CHON-001, was derived from the long bones of an 18-week female fetus.

Gene Expression: Positive for Collagen type I, Collagen type X, and aggrecan (RT-PCR).

Negative for Collagen type II (RT-PCR).

Antigen Expression: HLA-B58; Homo sapiens, expressed

HLA-B81; Homo sapiens, expressed

HLA-Cw6; Homo sapiens, expressed

HLA-Cw8; Homo sapiens, expressed

HLA-DR7; Homo sapiens, expressed

HLA-DR12; Homo sapiens, expressed

HLA-DR52; Homo sapiens, expressed

HLA-DQ2; Homo sapiens, expressed

HLA-DQ5; Homo sapiens, expressed

DNA Profile (STR): Amelogenin: X

CSF1PO: 10,11

D13S317: 12,14

D16S539: 11,13

D5S818: 11,12

D7S820: 12

THO1: 6,7

TPOX: 8,11

vWA: 16,17

Cytogenetic Analysis: CHON-001細(xì)胞This is a diploid cell line of female origin. Overall, the karyology is stable with a modal chromosome number of 46 in 93% of the examined cells and a low rate of polypoidy. No consistent structural chromosomal aberrations were found in any of the cells examined.

Age: 18 weeks fetus

Gender: female

Comments: The chondrocyte cell line, CHON-001, was derived from the long bones of an 18-week female fetus. The primary cells were infected by the defective retrovirus containing hTERT gene under G418 selection. The defective retrovirus was collected from the supernatant of the packaging cell line, PT67 transfected with the pLXSN vector containing hTERT gene. Gene Expression: Positive for Collagen type I, Collagen type X, and aggrecan (RT-PCR). Negative for Collagen type II (RT-PCR).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.

Propagation: ATCC complete growth medium: The base medium for this cell line is ATCC-formulated Dulbecco's Modified Eagle's Medium, Catalog No. 30-2002. To make the complete growth medium, add the following components to the base medium:

0.1mg/ml G-418

10% heat-inactivated fetal bovine serum

Atmosphere: air, 95%; carbon dioxide (CO2), 5%

Temperature: 37.0°C

Subculturing: Protocol: Volumes are given for T75 sq.cm flasks; proportionally reduce or increase the amount of dissociation medium for culture vessels of other sizes.

Remove and discard culture medium.

Add 2.0-3.0 ml of 0.05% Trypsin-0.53mM EDTA solution to the flask and observe cells under an inverted microscope until the cell layer is dispersed (usually within 5 to 10 minutes).

Note: CHON-001細(xì)胞To avoid clumping do not agitate the cells by hitting or shaking the flask while waiting for the cells to detach. Cells that are difficult to to detatch may be placed at 37C to facilitate dispersal.

Add 6.0 to 8.0 ml of complete growth medium and aspirate cells by gently pipetting.

Transfer cell suspension to a centrifuge tube and spin at approximately 125 x g for 5 to 15 minutes.

Discard supernatant and resuspend cells in fresh growth medium.

Add appropriate aliquots of cell suspension to new culture vessels. An inoculum of 2 X 10(3) to 3 X 10(3) viable cells/sq.cm is recommended. Subculture when cell concentration reaches between 1 x 10(4) and 2 x 10(4) cells/sq. cm.

Incubate cultures at 37 C.

Subcultivation Ratio: A subcultivation ratio of 1:4 to 1:6 is recommended

Preservation: Freeze medium: 90% heat-inactivated fetal bovine serum; 10% DMSO

Storage temperature: liquid nitrogen vapor phase

Doubling Time: about 72 hours

Related Products: Recommended medium (without the additional supplements or serum described under ATCC Medium):ATCC 30-2002

Cell culture tested DMSO:ATCC 4-X

Erythrosin B vital stain solution:ATCC 30-2404

Trypan Blue vital stain solution:ATCC 30-2402