LS-174T Cells
General information
Description | The line was derived from the same tumor as LS 180. LS 174T cells stain positively for cytokeratins. |
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Organism | Human |
Tissue | Colon |
Disease | Adenocarcinoma |
Synonyms | Ls174T, LS174t, Ls-174-T, LS-174-T, LS 174 T, LS174T, Ls-174T, LS 174T, LS-174, LS174 |
Characteristics
Age | 58 years |
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Gender | Female |
Ethnicity | Caucasian |
Morphology | Epithelial-like |
Growth properties | Adherent |
Identifiers / Biosafety / Citation
Citation | LS-174T (Cytion catalog number 300392) |
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Biosafety level | 1 |
Expression / Mutation
Protein expression | Colon Antigen 3 +, CEA +, p53 -, GFAP -, mRNA expression + |
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Antigen expression | HLA A2, B13, B50, Blood type O |
Isoenzymes | ADA, 1: G6PD, B, PGM1, 1, PGM3, 2, PGD, A, ES-D, 1, PEP-D, 1 |
Oncogenes | myc +, myb + , ras +, fos +, p53 +, sis -, abl -, ros -, src - |
Tumorigenic | Yes, in nude mice |
Reverse transcriptase | negative |
Products | Carcinoembryonic antigen (CEA) 1944 ng/106 cells in 10 days, mucin, interleukin-10 (IL-10), interleukin-6 (IL-6) |
Mutational profile | LS-174T cells carry a mutation in codon 12 of Kras gene: GGT(Wt Gly) >GAT(Asp) |
Karyotype | 45,x with one x chromosome missing but no other chromosomal aberrations |
Handling
Culture Medium | EMEM, w: 2 mM L-Glutamine, w: 1.5 g/L NaHCO3, w: EBSS, w: 1 mM Sodium pyruvate, w: NEAA (Cytion article number 820100c) |
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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 1:2 to 1:5 is recommended |
Seeding density | 5 to 8 x 10^4 cells/cm^2 |
Fluid renewal | 2 to 3 times per week |
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 |
Amelogenin: x,y
CSF1PO: 10,14
D13S317: 10,11
D16S539: 11,13
D5S818: 11,15
D7S820: 10.3,11
TH01: 6,7
TPOX: 8,9
vWA: 15,17,18,19
D3S1358: 15,17
D21S11: 29,30,31
D18S51: 11,13
Penta E: 15,16
Penta D: 10
D8S1179: 11,12,16
FGA: 21,22
D1S1656: 12,13,14,18.3,19.3
D6S1043: 12,13,14
D2S1338: 18,22
D12S391: 18,19,20
D19S433: 13,14,15
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HLA alleles |
A*: 02:xx, 30:01:01
B*: 13:xx, 35:01:01
C*: 04:01:01, 06:xx
DRB1*: 04:02:01, 07:01:01
DQA1*: 02:01:01, 03:01:01
DQB1*: 02:02:01, 03:02:01
DPB1*: 03:01:01G, 04:01:01
E: 01:01, 01:03
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