Immortalized Human Mammary Epithelial Progenitor (K5+/K19+) Cells - hTERT

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.

Immortalization Method: Serial passaging and transduction with recombinant retroviruses carrying hTERT gene

BioSafety Level: II

Species: Human

Source Organ: Mammary

Growth Properties: Adherent

Morphology: Cobble-stone|Spindle shaped

Population Doubling: N/A

Seeding Density: Thaw entire contents into an appropriate T25 flask as specified in the Propagation instructions.

Markers: K5, K14, vimentin, E-cadherin, p63, K8, K18, K19, CD29, CD49f, G418 resistance a-SMA, CD10, Thy-1 when grown in DCFI-1

Donor Age: N/A

Donor Gender: N/A

Donor Ethnicity: N/A

Propagation: 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 completed growth medium, add the following components to the base medium at final concentration: 0.01M HEPES (Gibco; 15630), fetal bovine serum (TM999)* to a final concentration of 1%, 12.5 ng/ml (EGF) epidermal growth factor (Z100135), 6.5 ng/ml triiodothyronine (Sigma; T5516), 0.545 ng/ml beta-estradiol (Sigma; E2257), 1 µg/ml insulin (Z101065), 1 µg/ml hydrocortisone (Sigma; I5500), 0.006X ethanolamine (Sigma; E9508), 14.1 µg/ml phosphoethanolamine (Sigma; P0503), 10 µg/ml transferrin (Sigma; T2252), 2 mM glutamine (Gibco; 25030-081), 2.6 ng/ml sodium selenite (Sigma; S1382), 1 ng/ml cholera toxin (Sigma; C8052), 35 µg/ml bovine pituitary extract (Hammond Cell Tech), Penicillin/Streptomycin Solution (G255) to a final concentration of 1% or 10 µg/ml Gentamicin (Gibco; 15750-060), and 10 µg/ml ascorbic acid (Sigma; A4034). Adjust to pH 7.4.
Carbon dioxide (CO₂): 5%, Temperature: +37.0°C.

* abm does not recommend to use heat-inactivated FBS for cell culture unless specified otherwise.

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

Shipping Condition: Dry Ice

Storage Condition: liquid nitrogen or -180°C