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HCE-2 [50.B1]

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價格:

編號:B161388

品牌:Mingzhoubio

標準菌株

定量菌液

DNA

RNA

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凍干粉

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甘油

平板

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產品名稱

HCE-2 [50.B1]

商品貨號

B161388

Organism

Homo sapiens, human

Tissue

eye, cornea

培養基

Cell Type

epithelial Adenovirus 12-SV40 hybrid transformed

Product Format

frozen

Morphology

epithelial

Culture Properties

adherent

Biosafety Level

2 [Cells contain Adeno-12/SV-40 viral 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.

Gender

male

Ethnicity

Black

Storage Conditions

liquid nitrogen vapor phase

Disclosure

This material is cited in a US or other Patent and may not be used to infringe the claims. Depending on the wishes of the Depositor, ATCC may be required to inform the Patent Depositor of the party to which the material was furnished. This material may not have been produced or characterized by ATCC.

Karyotype

aneuploid; Y chromosome is present

Images

Derivation

A primary culture of normal corneal epithelium was immortalized with Ad12-SV40 hybrid virus by overnight incubation with virus at 37°C

Clinical Data

Black

male

Effects

Yes, in soft agar

Comments

Keratinocyte Serum-Free medium is available from GIBCO, Grand Island, New York.

Complete Growth Medium

Keratinocyte-Serum Free Medium (Gibco 17005-042)
Supplemented with frozen additives included (from GIBCO):
1) 0.05 mg/ml bovine pituitary extract (BPE)
2) 5 ng/ml epidermal growth factor (EGF).
NOTE: Do not filter EGF
And also supplemented with 500 ng/ml hydrocortisone and 0.005mg/ml insulin (not included).

Subculturing

Volumes are given for a 75 cm² flask. Increase or decrease the amount of dissociation medium needed proportionally for culture vessels of other sizes.

Remove and discard culture medium.
Briefly rinse the cell layer with 0.05% (w/v) Trypsin-0.53mM EDTA solution.
Add 2.0 to 3.0 mL of Trypsin-EDTA solution to flask and observe cells under an inverted microscope until cell layer is dispersed (usually with 5 to 15 minutes).
Note: 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°C to facilitate dispersal.
Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting. (Or neutralize with medium containing 10% fetal bovine serum).
To remove trypsin-EDTA solution, transfer cell suspension to centrifuge tube and spin at approximately 125 x g for 5 to 10 minutes.
Discard supernatant and resuspend cells in fresh growth medium. Add appropriate aliquots of cell suspension to new culture vessels. Seed cells on flasks precoated with a mixture of 0.01 mg/ml fibronectin, 0.03 mg/mL bovine collagen type I and 0.01 mg/mL bovine serum albumin.
Incubate cultures at 37°C.

Subcultivation Ratio: A subcultivation ratio of 1:3 is recommended

Medium Renewal: Twice per week

Cryopreservation

Freeze medium: 85% Growth Medium, 10% FBS, 5% DMSO

Storage temperature: liquid nitrogen vapor phase

Culture Conditions

Temperature: 37°C

STR Profile

Amelogenin: X,Y

CSF1PO: 10,13

D13S317: 8,11

D16S539: 9,11

D5S818: 11,12

D7S820: 8,10

THO1: 6,9

TPOX: 10,11

vWA: 15,18

Isoenzymes

AK-1, 1

ES-D, 1

G6PD, A

GLO-I, 2

Me-2, 1-2

PGM1, 1

PGM3, 2

Population Doubling Time

30 hrs

Name of Depositor

CR Kahn, Gillette Medical Evaluation Laboratories

U.S. Patent Number

5,672,498

References

Kahn CR, Rhim J. Human corneal epithelial cell lines with extended lifespan. US Patent 5,585,265 dated Dec 17 1996

Walker TL, Kahn CR. Human corneal epithelial cell lines with extended lifespan. US Patent 5,672,498 dated Sep 30 1997

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