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CCD 1103 KIDTr
CCD 1103 KIDTr
規格:
價格:
編號:B164143
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
CCD 1103 KIDTr
商品貨號
B164143
Organism
Homo sapiens, human
Tissue
kidney
培養基
Cell Type
human papillomavirus 16 (HPV-16) E6/E7 transformed
Product Format
frozen
Morphology
epithelial
Culture Properties
adherent
Biosafety Level
2 [Cells contain Human Papilloma viral 16 (HPV16) 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.

Age
115 days gestation
Derivation
Fetal kidney was digested with collagenase: trypsin mixture. The digestion products were plated in REGM (Renal Epithelial Cell Growth Medium which includes 0.5% FBS) from Clonetics. Passage 3 cells were transformed with a retrovirus vector (LXSN16E6E7 produced by ATCC CRL-2203) in the presence of polybrene. The transformed cells are resistant to G418 and sensitive to Hygromycin B.
After 55 population doublings, the cells continue dividing and retain cuboidal morphology. E6E7 sequences were detected by PCR in cells at passage 15.
Oncogene
E6/E7 +
Genes Expressed
keratin; acid phosphatase
Cellular Products
keratin; acid phosphatase
Virus Susceptibility
Human Coxsackievirus B 5
Human Coxsackievirus A 9
Measles virus
Human poliovirus 1
Human respiratory syncytial virus
Comments
This line stains positively with antibody for cytokeratin and is acid phosphatase positive.

After 55 population doublings, the cells continue dividing and retain cuboidal morphology. E6E7 sequences were detected by PCR in cells at passage 15.

Major Histocompatibility Complex class I or II molecules were not expressed on these cells, but PCR analyses revealed presence of the genes for directing synthesis of HLA antigens.

CCD 1103 KIDTr is sensitive to: Coxsackievirus A9 and B5, measles virus, poliovirus 1 and respiratory syncytial virus (RSV).
Complete Growth Medium
The base medium for this cell line is ATCC-formulated F-12K Medium, Catalog No. 30-2004. To make the complete growth medium, add the following components to the base medium: fetal bovine serum to a final concentration of 10%.
Subculturing
Volumes used in this protocol are for 75 cm2 flask; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes. 
  1.  Remove and discard culture medium.
  2. Briefly rinse the cell layer with 0.25% (w/v) Trypsin-0.53mM  EDTA solution to remove all traces of serum which contains trypsin inhibitor.
  3. 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 within 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.
  4. Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting. 
  5. Add appropriate aliquots of the cell supension to new culture vessels.
  6. Incubate cultures at 37°C.
Subcultivation Ratio: 1:3 to 1:5.

Medium Renewal: Two to three times weekly.

Note: For more information on enzymatic dissociation and subculturing of cell lines consult Chapter 10 in Culture of Animal Cells, a manual of Basic Technique by R. Ian Freshney, 3rd edition, published by Alan R. Liss, N.Y., 1994.   

Cryopreservation
culture medium 95%; DMSO, 5%
Culture Conditions
Temperature: 37°C
Name of Depositor
A Thompson
References

Zabrenetzky V, et al. The isolation, immortalization and characterization of human fetal keratinocytes. In Vitro Cell. Dev. Biol. 33: Part II, p. 34A, 1997.

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