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HT29-MTX resistant (10 microMolar)

HT29-MTX resistant (10 microMolar)

Catalogue No.

15121711

Cell Line Name

HT29-MTX resistant (10 microMolar)

Cell Line Description

HT29-MTX resistant (10 microMolar) is a cell population isolated from the human colon cancer cell line HT29 by adaption to 10 µM Methotrexate (MTX). Methotrexate interferes with a metabolic pathway, namely that of folate, by inhibiting the enzyme Dihydrofolate reductase (DHFR). HT29-MTX-resistant cells form a homogenous population of polarised goblet cells that secrete mucins of gastric immunoreactivity. A mucus gel becomes visible on the cell surface of the cell monolayer at post-confluency. These cells maintain their ability to differentiate under normal culture conditions in the absence of methotrexate. This cell line is tumourgenic in nude mice, with xenografts from MTX-adapted cells well differentiated and strongly express both mucus and DPP-IV (Dipeptidyl Peptidase IV). All cells show an apical expression of DPP-IV, CEA and villin regardless of whether they secrete mucins. P-glycoprotein was undetected in this cell line according to immunofluorescence staining.

General Info

Species

Human

Unique to ECACC

Yes

Also Known As

HT29MTX, HT29MTX resistant, HT29-MTX resistant, HT29-MTX resistant (10-5M)

Characteristics

Products

Cells form a homogenous population of polarised goblet cells that secrete mucins with gastric immunoreactivity.

Tissue of Origin

Colon

Morphology

Epithelial

DNA profile (STR Profile)

Amelogenin: X

CSF1PO: 11, 12

D3S1358: 15, 17

D5S818: 11, 12

D7S820: 9.3

D8S1179: 10

D13S317: 11

D16S539: 11, 12

D18S51: 13, 20.2

D21S11: 29

FGA: 20, 22

Penta D: 11

Penta E: 14, 16

TH01: 6, 9

TPOX: 8, 9

vWA: 17

Applications

Defining the biological characteristics of therapeutic drug resistance in tumour cells to develop new treatment strategies

Disease

Cancer

Culture Conditions

Cell Type

Epithelial

Subculture Routine

Split sub-confluent cultures (70-80%) 1:3 to 1:6. Seeding can be as low as 0.32 x 10⁴ cells/cm² up to 4 x 10⁴ cells/cm², using 0.05% trypsin/EDTA solution. Cells are cultured at 37°C, in 90% air and 10% CO₂, and passaged every 3 to 4 days.

Culture Medium

DMEM with 10% heat-inactivated Foetal Bovine Serum (FBS) and 10µM Methotrexate (MTX). Cryopreserve cells in 10% Dimethyl sulphoxide (DMSO) and 90% FBS.

Growth Mode

Adherent

Additional Info

Depositor

CelluloNet, France on behalf of Thecla Lesuffleur (INSERM)

Country of Origin

France

Hazard Group (ACDP)

Hazard Group (ACDP) 2

Applications

References

Leteurtre E, Gouyer V, Rousseau K, Moreau O, Barbat A, Swallow D, Huet G, Lesuffleur T. Differential mucin expression in colon carcinoma HT-29 clones with variable resistance to 5-fluorouracil and methotrexate. Biol Cell. 2004 Mar;96 (2):145-51. PMID: 15050369 Lesuffleur T, Violette S, Vasile-Pandrea I, Dussaulx E, Barbat A, Muleris M, Zweibaum A. Resistance to high concentrations of methotrexate and 5-fluorouracil of differentiated HT-29 colon-cancer cells is restricted to cells of enterocytic phenotype. Int J Cancer. 1998 May 4;76 (3):383-92. PMID: 9579576

Bibliography

Lesuffleur T, Porchet N, Aubert JP, Swallow D, Gum JR, Kim YS, Real FX, Zweibaum A. Differential expression of the human mucin genes MUC1 to MUC5 in relation to growth and differentiation of different mucus-secreting HT-29 cell subpopulations. J Cell Sci. 1993 Nov;106 (Pt 3):771-83. PMID: 8308060.

Available Formats

  • Frozen
  • DNA-5µg (100ng/µl)

If use of this culture results in a scientific publication, it should be cited in the publication as: HT29-MTX resistant (10 microMolar) (ECACC 15121711).

Unless specified otherwise, at ECACC we routinely handle all of our cell lines at containment level 2 in accordance with the ACDP guidelines (Advisory Committee on Dangerous Pathogens) (UK). All cell cultures have the potential to carry as yet unidentified adventitious agents. It is the responsibility of the end user to ensure that their facilities comply with biosafety regulations for their own country. ACDP Guidance: Biological agents: Managing the risks in laboratories and healthcare premises.

The Culture Collections represent deposits of cultures from world-wide sources. While every effort is made to ensure details distributed by Culture Collections are accurate, Culture Collections cannot be held responsible for any inaccuracies in the data supplied. References where quoted are mainly attributed to the establishment of the cell culture and not for any specific property of the cell line, therefore further references should be obtained regarding cell culture characteristics. Passage numbers where given act only as a guide and Culture Collections does not guarantee the passage number stated will be the passage number received by the customer.

Cultures supplied by Culture Collections are for research purposes only. Enquiries regarding the commercial use of a cell line are referred to the depositor of the cell line. Some cell lines have additional special release conditions such as the requirement for a material transfer agreement to be completed by the potential recipient prior to the supply of the cell line. Please view the Terms & Conditions of Supply for more information.