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Immortalized Human Mammary Epithelial Cells (HMEC 2.6) - SV40

SKU : BHC10900584

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Applied Biological Materials (abm) Inc.

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Review(s)

Mfr.No.	T0455
Description	The immortalized human mammary progenitor cells hTERT (K5+/K19+) is a clonal cell population of progenitors coexpressing normal mammary and stem cell markers. It has the ability to self-renew and differentiate into luminal and/or myoepithelial cell lineages. Through propagation in different optimized media, the cells are able to give rise to new population of cells with different morphology and characteristics, such as mucin-1 positive cells, vimentin-negative cells, or expressing basal, luminal, and other stem cell markers. Aldehyde dehydrogenase 1A3 enzyme is noticeably higher in expression, a marker commonly used for isolating normal and tumor mammary stem cells. K5+/K19+ cells have Wnt, Notch, and hedeghog gene regulatory pathways. It is a valuable tool in research in the field of biology, the stem/progenitor origin, and the heterogeneity mechanisms in breast cancer.
Cell Type	Immortalized Cells
Disclaimer	1. For for-profit organizations and corporations, please contact [email protected] for pricing of this item. 2. Sale of this item is subjected to the completion of a Material Transfer Agreement (MTA) by the purchasing individual/institution for each order. If you have any questions regarding this, please contact us at [email protected]. 3. All test parameters provided in the CoA are conducted using abm's standardized culture system and procedures. The stated values may vary under the end-user's culture conditions. Please verify that the product is suitable for your studies by referencing published papers or ordering RNA (0.5 mug, Cat.# C207, $450.00) or cell lysate (100 mug, Cat.# C206, $600.00) to perform preliminary experiments, or alternatively use our Gene Expression Assay Service (Cat# C138). All sales are final. 4. We recommend live cell shipments for ease of cell transfer and this option can be requested at the time of ordering. Please note that the end-user will need to evaluate the feasibility of live cell shipment by taking into account the final destination's temperature variation and its geographical location. In addition, we thoroughly test our cell lines for freeze-thaw recovery. If frozen cells were received and not recovered in your lab under the exact, specified conditions (using recommended culture vessel, media, additional supplements, and atmospheric conditions), a live cell replacement is possible at a cost (plus shipping). 5. All of abm's cell biology products are for research use ONLY and NOT for therapeutic/diagnostic applications. abm is not liable for any repercussions arising from the use of its cell biology product(s) in therapeutic/diagnostic application(s). Please contact a technical service representative for more information. 6. abm makes no warranties or representations as to the accuracy of the information on this site. Citations from literature and provided for informational purposes only. abm does not warrant that such information has been shown to be accurate. 7. abm warrants that cell lines shall be viable upon initiation of culture for a period of thirty (30) days after shipment and that they shall meet the specifications on the applicable abm Material Product Information sheet, certificate of analysis, and/or catalog description. Such thirty (30) day period is referred to herein as the "Warranty Period."
Expression Level	K5, K14, vimentin, E-cadherin, p63, K8, K18, K19, CD29, CD49f a-SMA, CD10, Thy-1 when grown in DCFI-1
Quantity	1x106 cells / 1.0 ml
Tissue	Breast
Morphology	Cobble-stone\|Spindle shaped
Population Doubling	24 - 48 hours
Species	Human (H. sapiens)
Propagation	Use of PriCoatTM T25 Flasks (G299) or Applied Cell Extracellular Matrix (G422) is required for cell adhesion to the culture vessels. Grow cells in ECM-coated culture vessels unless otherwise specified in the Propagation Requirements below.Optimized Media:To express basal, luminal, and stem cell markers with some myoepithelial cell markers (a-SMA, CD10, Thy-1), grow cells in DFCI-1 media.To make DFCI-1 the base medium is Prigrow VIII and Prigrow IV (1:1, vol/vol) medium available at abm (TM018 and TM004). To make the complete growth medium, add the following components to the base medium: fetal bovine serum (TM999)* to a final concentration of 1%, Penicillin/Streptomycin Solution (G255) to a final concentration of 1%, HEPES (Gibco) to a final concentration of 10 mM, 12.5 ng/ml Recombinant Human EGF (Z100135) to a final concentration of 12.5 ng/ml, 6.5 ng/ml triiodothyronine (Sigma), 0.545 ng/ml beta-estradiol (Sigma), 1 µg/ml insulin (TM053), 1 µg/ml hydrocortisone (Sigma), 0.006X ethanolamine (Sigma), 14.1 µg/ml phosphoethanolamine (Sigma), 10 µg/ml transferrin (Sigma), 2 mM L-glutamine (G275), 2.6 ng/ml sodium selenite (Sigma), 1 ng/ml cholera toxin (Sigma), 35 µg/ml bovine pituitary extract (Hammond Cell Tech), and 10 µg/ml ascorbic acid (Sigma). Adjust pH to 7.4.Change media every 2-3 days.Carbon dioxide (CO2): 5%, Temperature: 37.0°C.* Do not use heat-inactivated FBS for cell culture unless specified otherwise.For cells to adopt spindle-shape morphology around tight epithelial cell colonies and for the presence of mucin-1 positive cells: grow cells in mammary epithelial growth medium (MEGM; Lonza) supplemented with B27 (10ml/500ml medium), 20 ng/ml Recombinant Human EGF (Z100135), 20 ng/ml Recombinant Human FGF2 (Z101455), 4 µg/ml heparin, and Penicillin/Streptomycin Solution (G255) to a final concentration of 1%.Carbon dioxide (CO2): 6.5%, Temperature: 37.0°C.For luminal differentiation, the absence of myoepithelial differentiation, and the appearance of mucin-1 positive and vimentin-negative cells: grow the cells in DCFI-2 media where the base is Prigrow VIII and Prigrow IV (1:1, vol/vol) medium available at abm (TM018 and TM004). Supplement with 10 mM HEPES (Gibco), 12.5 ng/ml Recombinant Human EGF (Z100135), 6.5 ng/ml triiodothyronine (Sigma), 0.545 ng/ml beta-estradiol (Sigma), 1 µg/ml insulin (TM053), 1 µg/ml hydrocortisone (Sigma), 0.006X ethanolamine (Sigma), 14.1 µg/ml phosphoethanolamine (Sigma), 10 µg/ml transferrin (Sigma), 2 mM L-glutamine (G275), 2.6 ng/ml sodium selenite (Sigma), 1 ng/mg cholera toxin (Sigma), 0.05% bovine serum albumin (BSA), Penicillin/Streptomycin Solution (G255) to a final concentration of 1%, and 10 µg/ml ascorbic acid (Sigma). Adjust pH to 7.4.Carbon dioxide (CO2): 6.5%, Temperature: 37.0°C.
Quality Control	1) Western blot to assess cell lineage-related and stem cell markers; 2) Immunofluorescence staining analysis for lineage markers; 3) Gene expression profiling for gene regulatory pathways

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SKU	Supplier No.	Your price	Quantity
BHC10900584	T0455	$1085		Add to Cart

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