LS513 (ATCC® CRL-2134)

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
Biosafety Level 1
Disease Dukes' type C,colorectal carcinoma
Age 63 years
Gender male
Ethnicity Caucasian
Storage Conditions liquid nitrogen vapor phase
Karyotype Two stem lines could be distinguished. The main one was represented in 65% of the cells, with a modal number of 51,XY and 3 markers, M1 - der(1)t(1;15), M2 - der(2)t(2;3)der(3)t(2;3), M3, and a monosomy 15., The second stem line had a modal chromosome number of 52,XY and presented M2 and M3 plus an isochromosome for the long arm of chromosome 1 called M4., A trisomy 5,7, a tetrasomy 13, and a monosomy 2 and 3 were present in all of the cells analyzed; the line did not exhibit monosomy 15.
Derivation
LS513 is a colorectal carcinoma cell line isolated in 1985 from a primary tumor biopsy from a 63 year old Caucasian male patient diagnosed with a Dukes' C mucin secreting cecal tumor located at the Bauhin valve.
Clinical Data
63 years
Caucasian
male
Antigen Expression
carcinoembryonic antigen (CEA); ICAM-1; HLA class I positive
Oncogene p53 + (wild type)
Genes Expressed
transforming growth factor beta 1 (TGF beta-1, 83 pg per 106 cells per 24 hours)
Tumorigenic Yes
Effects
Yes, forms tumors in nude mice
Comments
LS513 is a colorectal carcinoma cell line isolated in 1985 from a primary tumor biopsy from a 63 year old Caucasian male patient diagnosed with a Dukes' C mucin secreting cecal tumor located at the Bauhin valve.

Approximately 50% of LS513 cells express surface carcinoembryonic antigen (CEA).

LS513 cells express the major histocompatibility (MHC) class I antigens HLA and beta 2 microglobulin.

MHC class II antigens (HLA-DR, DQ, and DP were not detected).

TGF beta-1 is inhibitory for proliferation of LS513 cells, whereas TGF beta-2 has no effect on the growth of these cells.

LS513 cells are 100-fold less sensitive to TGF beta-1 than the LS1034 (ATCC CRL-2158) cell line.

LS513 cells are multidrug resistant (MDR) and are tumorigenic in nude mice.

Colony forming efficiency was 30% in methylcellulose.



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 to 1:4
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: 50% Culture Medium + 40% FBS + 10% DMSO
Storage temperature: liquid nitrogen vapor phase
STR Profile
Amelogenin: X,Y
CSF1PO: 10,13
D13S317: 9,10
D16S539: 12,13
D5S818: 9,11
D7S820: 8,11
THO1: 8
TPOX: 8
vWA: 16,17
Population Doubling Time 25 hrs
Name of Depositor L Suardet
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

Basic 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