hTERT RPE-1細(xì)胞

器官來源： 其他

生長狀態(tài)： 貼壁生長

細(xì)胞類型： 其他細(xì)胞類型

數(shù)量： 大量

細(xì)胞形態(tài)： 上皮樣

是否是腫瘤細(xì)胞： 0

物種來源： 人

運輸方式： 凍存運輸

ATCC Number： CRL-4000?

組織來源： epithelium, pigmented

Designations: hTERT RPE-1

Depositors: Geron Corporation

hTERT RPE-1細(xì)胞Biosafety Level: 1

Shipped: frozen

Medium & Serum: See Propagation

Growth Properties: adherent

Organism: Homo sapiens

Morphology: epithelial

Source: Organ: retina, eye

Tissue: epithelium, pigmented

Cell Type: epithelial 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. hTERT RPE-1細(xì)胞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.

Restrictions: This material requires that the Addendum for Commercial and For-Profit Organizations or the Addendum for Noncommercial and Academic Organizations be signed and returned to ATCC before shipment. The price listed above is for noncommercial and academic organizations only. Commercial and for-profit organizations should call for pricing.

Applications: s 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 hTERT-immortalized retinal pigment epithelial cell line, hTERT RPE-1, was derived by transfecting the RPE-340 cell line with the pGRN145 hTERT-expressing plasmid (ATCC MBA-141).

Antigen Expression: Ep-16 epithelial antigen; Homo sapiens, expressed (The Ep-16 antigen was assayed by flow cytometry using the Ep-16 monoclonal antibody (ATCC HB-155)) (The Ep-16 antigen was assayed by flow cytometry using the Ep-16 monoclonal antibody (ATCC HB-155))

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

DNA Profile (STR): hTERT RPE-1細(xì)胞Amelogenin: X

CSF1PO: 12,14

D13S317: 11,12

D16S539: 11

D5S818: 11

D7S820: 10,11

THO1: 9

TPOX: 8

vWA: 17,18

Cytogenetic Analysis: This is a near-diploid human cell line of female origin with a modal chromosome number of 46 that occurred in 90% of the cells counted. The sex chromosomes consist of a karyotypically normal X-chromosome and a derivative X-chromosome with additional chromosomal material at the terminal end of the q-arm. The derivative X-chromosome was present in all of the cells analyzed.

Gender: female

Comments: The hTERT-immortalized retinal pigment epithelial cell line, hTERT RPE-1, was derived by transfecting the RPE-340 cell line with the pGRN145 hTERT-expressing plasmid (ATCC MBA-141). Cells were cultured in medium containing hygromycin B until stable clones were selected [Pubmed: 9454332, 9916802].

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. hTERT RPE-1細(xì)胞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 DMEM:F12 Medium Catalog No. 30-2006.To make the complete growth medium, add the following components to the base medium:

fetal bovine serum to a final concentration of 10%

0.01 mg/ml hygromycin B

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

Temperature: 37.0°C

Growth Conditions: Subculture when cell concentration reaches between 2 X 10(4) and 4 X 10(4) cells/cm2.

Subculturing: Protocol: Volumes used in this protocol are for 75 cm2 flasks; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes.

Remove and discard culture medium.

Briefly rinse the cell layer with Hanks? Balanced Salt Solution (HBSS)or 0.25% (w/v) Trypsin - 0.53 mM EDTA solution to remove all traces of serum which contains trypsin inhibitor.

Add 3.0 to 5.0 ml of Trypsin-EDTA solution to flask and observe cells under an inverted microscope until cell layer is dispersed (usually within 5 to 15 minutes).

Note: hTERT RPE-1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 detach may be placed at 37.0°C 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 4 X 103 to 6 X 103 viable cells/cm2 is recommended.

Incubate cultures at 37.0°C.

Subcultivation Ratio: 1:5 to 1:10 twice weekly

Medium Renewal: every 2 days (or as needed)

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

Storage temperature: liquid nitrogen vapor phase

Doubling Time: 19 hr

Related Products: Recommended medium (without the additional supplements or serum described under ATCC Medium):ATCC 30-2006

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

Hanks' Balanced Salt Solution (HBSS): ATCC 30-2213

References: 47354: 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

90422: Matsunaga H, et al. Beta-galactosidase histochemistry and telomere loss in senescent retinal pigment epithelial cells. Invest. Ophthalmol. Vis. Sci. 40: 197-202, 1999. PubMed: 9888444