Immortalized Human Mammary Epithelial Progenitor (K5+/K19+) Cells - hTERT | T0452

Immortalized Human Mammary Epithelial Progenitor (K5+/K19+) Cells - hTERT | T0452

+/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

Organism:

Homo sapiens

Species:

Human

Source Organ:

Mammary

Organ Type:

Breast

Growth Properties:

Adherent

Morphology:

Cobble-stone|Spindle shaped

Passage Number:

N/A

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 . 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 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 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.

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 -180C