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UK Health Security Agency

HMT-3522 S2

HMT-3522 S2

Catalogue No.

98102211

Cell Line Name

HMT-3522 S2

Cell Line Description

The cell line HMT-3522 S2, also known as HMT-3522/gt1, is a subline that has been derived from HMT-3522. The parent line was established from a benign breast tumour of a 48 year old woman and has undergone spontaneous malignant transformation. It was grown on collagen in serum-free media without antibiotics from explantation onwards. A mutation at codon 179 of the p53 gene was detected during establishment of the parent line at passage 25-30. The subline S1 is a continuation of this line from passage 34 (ECACC catalogue no. 98102210) growing without collagen. Subline S2 has been isolated by culturing S1 cells from passage 118 in serum-free media without epidermal growth factor (EGF). A third subline, T4-2 (ECACC catalogue no. 98102212) was established from S2 at passage 238 and is the only tumorigenic line out of the 3 sublines. S2 and T4-2 grow on collagen-coated flasks. HMT-3522 S2 is near-diploid, but an evolution of karyotype has been observed by regular chromosomal analyses. The expression of cytokeratin 13, 14, 17 and 18 and vimentin has been reported. Overexpression of mRNAs for EGF-R, TGF-alpha, TGF-beta and c-erb-B2 compared to subline S1 has been shown. In addition, expression of p53 mRNA has been reported. The cells have been tested positive for EGF receptor but negative for estrogen receptor. Growth is inhibited by addition of EGF. The decreased growth factor requirements developed simultaneously with amplification and overexpression of c-myc protooncogene. Publications using this cell line must refer to the original paper (Briand et al., In Vitro Cell Dev Biol 1987; 23:181). Any sublines created must include the name €œHMT-3522€ in the designation, but a short name may be used in the same paper. Further references are available on request.

Characteristics

Receptors

EGF

Tissue of Origin

Breast

DNA profile (STR Profile)

Amelogenin: X
CSF1PO: 11,12
D5S818: 9,11
D7S820: 9
D13S317: 11
D16S539: 11,13
TH01: 9,9.3
TPOX: 11
vWA: 14,17

Karyotype

42,XX,1p-,3p-,-6,7q-,8q-,8p-,9p+,12p+,-13,14q+,15p+,17q+,-18,19q+,-20,21p+

Applications

Evolution of karyotype and growth factor requirements, model for premalignant breast epithelium and study of mechanism of malignant conversion

Disease

Carcinoma

Culture Conditions

Cell Type

Epithelial-like

Subculture Routine

If starting from a frozen ampoule the cryoprotectant should be removed. Add thawed cells to a conical based centrifuge tube e.g. 15ml tube; slowly add 4 ml of culture medium to the tube. Take a sample of the cell suspension, e.g. 100µl, to count cells. Centrifuge the cell suspension at low speed i.e. 100 - 150 x g for a maximum of 5 minutes.Split subconfluent cultures using 0.05% trypsin or trypsin/EDTA; 5% CO₂; 37°C. Seed pre-coated flasks at 2-4 x10, 000 cells/cm². Cells detach slowly; add same volume of trypsin inhibitor as trypsin used, centrifuge and reseed cells in collagen IV coated flasks. Medium is serum-free, therefore trypsin inhibitor is essential. Cells can be slow in spreading out after subculture and may take 2-3 days to attach. Good growth occurs 5-7 days after subculture.Cells should be frozen in (45% conditioned media: 45% fresh media) with 10% DMSO.

Culture Medium

DMEM / Ham's F12 (1:1) + 2mM Glutamine + 250ng/ml insulin + 10µg / ml transferrin +10⁻⁸M Sodium selenite + 10⁻¹⁰M 17 β-estradiol + 0.5µg/ml hydrocortisone + 5µg / ml ovine prolactin

Growth Mode

Adherent

Additional Info

Depositor

Dr P Briand, Department of Tumour Endocrinology, Institute of Cancer Biology, Danish Cancer Society, Copenhagen, Denmark

Country of Origin

Denmark

Hazard Group (ACDP)

Hazard Group (ACDP) 2

Applications

References

In Vitro Cell Dev Biol 1987; 23:181;Cancer Res 1992;52:1210; Cancer Res 1996;56:2039; J Cell Biol 1997; 137: 231; Genes Chromosom Cancer 1997; 20: 30; Nat Biotechnol 1997; 15: 517

Available Formats

  • Frozen

If use of this culture results in a scientific publication, it should be cited in the publication as: HMT-3522 S2 (ECACC 98102211).

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.