Panc 04.03細(xì)胞,， 人胰腺癌細(xì)胞

Panc 04.03 (ATCC® CRL-2555™)

Organism  Homo sapiens, human

Tissue  pancreas

Product Format  frozen

Morphology  epithelial

Culture Properties  adherent

Biosafety Level  1

Disease  adenocarcinoma

Age  70 years adult

Gender  male

Ethnicity  White

Storage Conditions  liquid nitrogen vapor phase

Derivation  Panc 04.03 is a pancreatic adenocarcinoma epithelial cell line derived, in 1995, from a primary tumor removed from the head-of-the-pancreas of a male with pancreatic adenocarcinoma.

Clinical Data  70 years

Caucasian

male

Antigen Expression  MHC class I +; MHC class II -

RefJaffee EM, et al. Development and characterization of a cytokine-secreting pancreatic adenocarcinoma vaccine from primary tumors for use in clinical trials. Cancer J. Sci. Am. 4: 194-203, 1998. PubMed: 9612602

Blood type O; Rh+

Oncogene  K-ras + RefJaffee EM, et al. Development and characterization of a cytokine-secreting pancreatic adenocarcinoma vaccine from primary tumors for use in clinical trials. Cancer J. Sci. Am. 4: 194-203, 1998. PubMed: 9612602

Genes Expressed  cytokeratins 7 and 18 RefJaffee EM, et al. Development and characterization of a cytokine-secreting pancreatic adenocarcinoma vaccine from primary tumors for use in clinical trials. Cancer J. Sci. Am. 4: 194-203, 1998. PubMed: 9612602

Tumorigenic  Yes

Effects  Yes, forms tumors in nude or SCID mice

Comments  The cell line exhibits a K-ras oncogene mutation at codon 12 where a GGT --> GAT mutation resulted in substitution of aspartic acid for glycine. RefJaffee EM, et al. Development and characterization of a cytokine-secreting pancreatic adenocarcinoma vaccine from primary tumors for use in clinical trials. Cancer J. Sci. Am. 4: 194-203, 1998. PubMed: 9612602

The cells have a reported plating efficiency of 80%. Ref

Panc 04.03細(xì)胞， 人胰腺癌細(xì)胞

Complete Growth Medium  The base medium for this cell line is ATCC-formulated RPMI-1640 Medium, Catalog No. 30-2001. To make the complete growth medium, add the following components to the base medium:

•20 Units/ml human recombinant insulin

•fetal bovine serum to a final concentration of 15%

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-053mM 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 suspension to new culture vessels.

6.Incubate cultures at 37°C.

Subc*tion Ratio: 1:2 to 1:3

Medium Renewal: Add media once per week. Fluid change one to two times per week.

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  Complete growth medium supplemented with 5% (v/v) DMSO. Cell culture tested DMSO is available as ATCC Catalog No. 4-X.

Culture Conditions  Temperature: 37°C