LS-513 Cells
General information
Description | The colorectal carcinoma cell line LS-513 was isolated in 1985 from a primary tumor biopsy of a 63 year old Caucasian male patient. He was diagnosed with a Dukes' C mucin secreting cecal tumor located at the Bauhin valve. LS-513 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 LS-513 cells, whereas TGF beta-2 has no effect on the growth of these cells. LS-513 cells are 100-fold less sensitive to TGF beta-1 than the LS-1034 cell line. LS-513 cells are multidrug resistant (MDR) and are tumorigenic in nude mice. Colony forming efficiency was 30% in methylcellulose. |
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Organism | Human |
Tissue | Colorectal |
Disease | Adenocarcinoma |
Synonyms | LS513, LS 513 |
Characteristics
Age | 63 years |
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Gender | Male |
Ethnicity | Caucasian |
Morphology | Epithelial-like |
Growth properties | Adherent |
Identifiers / Biosafety / Citation
Citation | LS-513 (Cytion catalog number 300457) |
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Biosafety level | 1 |
Expression / Mutation
Protein expression | CEA+ (50%), p53+ |
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Antigen expression | Carcinoembryonic antigen (CEA), ICAM-1, HLA class I positive |
Tumorigenic | Yes, forms tumors in nude mice |
Products | Transforming growth factor beta 1 (TGF beta-1, 83 pg per 10 exp6 cells per 24 hours) |
Karyotype | Two stem lines can 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. |
Handling
Culture Medium | Ham's F12, w: 1.0 mM stable Glutamine, w: 1.0 mM Sodium pyruvate, w: 1.1 g/L NaHCO3 (Cytion article number 820600a) |
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Medium supplements | Supplement the medium with 10% FBS |
Passaging solution | Accutase |
Subculturing | Remove the old medium from the adherent cells and wash them with PBS that lacks calcium and magnesium. For T25 flasks, use 3-5 ml of PBS, and for T75 flasks, use 5-10 ml. Then, cover the cells completely with Accutase, using 1-2 ml for T25 flasks and 2.5 ml for T75 flasks. Let the cells incubate at room temperature for 8-10 minutes to detach them. After incubation, gently mix the cells with 10 ml of medium to resuspend them, then centrifuge at 300xg for 3 minutes. Discard the supernatant, resuspend the cells in fresh medium, and transfer them into new flasks that already contain fresh medium. |
Split ratio | A ratio of 1:3 to 1:4 is recommended |
Seeding density | 1 x 10^4 cells/cm^2 |
Fluid renewal | Every 3 days |
Freezing recovery | After thawing, plate the cells at 5 x 10^4 cells/cm^2 and allow the cells to recover from the freezing process and to adhere for at least 24 hours. |
Freeze medium | CM-1 (Cytion catalog number 800100) or CM-ACF (Cytion catalog number 806100) |
Handling of cryopreserved cultures |
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Quality control / Genetic profile / HLA
Sterility | Mycoplasma contamination is excluded using both PCR-based assays and luminescence-based mycoplasma detection methods. To ensure there is no bacterial, fungal, or yeast contamination, cell cultures are subjected to daily visual inspections. |
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STR profile |
CSF1PO: 10
D13S317: 9,10
D16S539: 12,13
D5S818: 11
D7S820: 8,11
TH01: 8
TPOX: 8
vWA: 16,17
D3S1358: 15
D21S11: 30
D18S51: 12,18
Penta E: 5,18
Penta D: 9,14
D8S1179: 13
FGA: 19,21
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HLA alleles |
A*: 32:01:01
B*: 51:01:01
C*: 01:02:01
DRB1*: 11:01:01
DQA1*: 05:05:01
DQB1*: 03:01:01
DPB1*: 04:01:01
E: 01:01:01
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