LS1034 (ATCC® CRL-2158)

Organism: Homo sapiens, human  /  Tissue: Cecum  /  Disease: Dukes' type C, colorectal carcinoma

Organism Homo sapiens, human
Tissue Cecum
Product Format frozen
Morphology epithelial
Culture Properties adherent, adherent
Biosafety Level 1
Disease Dukes' type C, colorectal carcinoma
Age 54 years
Gender male
Ethnicity Caucasian
Storage Conditions liquid nitrogen vapor phase
Karyotype modal number = 77, YY; seven markers involving chromosomes 1, 7, 14, X, and 18 were recognized. The other chromosomes involved could not be properly identified.; Monosomy 7 and monosomy 14 were present in all 15 metaphases analyzed. No normal chromosomes 5 or 18 were observed.
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Derivation
LS1034 is a colorectal carcinoma cell line isolated in 1989 from a primary tumor biopsy from a 54-year old Caucasian male patient diagnosed with Dukes' Type C, moderately to poorly differentiated cecal carcinoma.
Clinical Data
54 years
Caucasian
male
Antigen Expression
carcinoembryonic antigen (CEA); ICAM-1; HLA class I positive
Oncogene p53 + (mutated, Gly --> Ser mutation at position 245), APC (mutated, deletion, GAAAAGATT --> GATT at codon 1309)
Tumorigenic Yes
Effects
Yes, forms tumors in nude mice
Comments
More than 90% of LS1034 cells express surface CEA.
The cells express the major histocompatibility (MHC) class I antigens and beta 2 microglobulin, but class II antigens, (HLA-DR, DQ, and DP) were not detected.
No measurable amount of latent transforming growth factor beta-1 (TGF beta-1) is secreted.
Picomolar concentrations of TGF beta-1, TGF beta-2, and TGF beta-3 inhibit the proliferation of LS1034 cells.
LS1034 cells are multidrug resistant (MDR), and can proliferate in serum free medium.
The patient did not exhibit familial adenomatous polyposis (FAP) and normal tissue did not have the deletion in the APC gene.
The colony forming efficiency was 3% in methylcellulose medium containing 5% fetal bovine serum, and the clonogenicity is increased 600% in serum free methylcellulose as compared to semisolid medium containing serum.
A culture submitted to the ATCC in September 1994 was found to be contaminated with mycoplasma, and was cured by a 21 day treatment with BM Cycline.
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: 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-0.53 mM EDTA solution to remove all traces of serum that 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

Subculture Ratio: 1:3
Medium Renewal: 2 to 3 times a 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
Freeze Medium: Complete growth medium supplemented with 5% (v/v) DMSO
Storage Temperature: Liquid nitrogen vapor phase
Culture Conditions
Temperature: 37°C
STR Profile
Amelogenin: X,Y
CSF1PO: 11,12
D13S317: 12
D16S539: 9,11
D5S818: 12,13
D7S820: 8,12
THO1: 7
TPOX: 9,11
vWA: 17
Population Doubling Time 33 hrs
Name of Depositor L Suardet
Year of Origin 1989
References

Suardet L, et al. Responsiveness of three newly established human colorectal cancer cell lines to transforming growth factors beta 1 and beta 2. Cancer Res. 52: 3705-3712, 1992. PubMed: 1617643

Burmester JK, et al. Characterization of distinct functional domains of transforming growth factor beta. Proc. Natl. Acad. Sci. USA 90: 8628-8632, 1993. PubMed: 7690965

Li CY, et al. Potential role of WAF1/Cip1/p21 as a mediator of TGF-beta cytoinhibitory effect. J. Biol. Chem. 270: 4971-4974, 1995. PubMed: 7890601

Cottrell S, et al. Molecular analysis of APC mutations in familial adenomatous polyposis and sporadic colon carcinomas. Lancet 340: 626-630, 1992. PubMed: 1355210

Basic Documentation
Other Documentation
References

Suardet L, et al. Responsiveness of three newly established human colorectal cancer cell lines to transforming growth factors beta 1 and beta 2. Cancer Res. 52: 3705-3712, 1992. PubMed: 1617643

Burmester JK, et al. Characterization of distinct functional domains of transforming growth factor beta. Proc. Natl. Acad. Sci. USA 90: 8628-8632, 1993. PubMed: 7690965

Li CY, et al. Potential role of WAF1/Cip1/p21 as a mediator of TGF-beta cytoinhibitory effect. J. Biol. Chem. 270: 4971-4974, 1995. PubMed: 7890601

Cottrell S, et al. Molecular analysis of APC mutations in familial adenomatous polyposis and sporadic colon carcinomas. Lancet 340: 626-630, 1992. PubMed: 1355210