Immortalized Human Mammary Fibroblasts (HMF3S) were generated through the combined transduction of mammary microvascular endothelia cells with a temperature sensitive mutant (U19tsA58) of the SV40 Large T Antigen and a catalytic subunit of human telomerase (hTERT). The HMF3S cells grow best at a temperature of +37⁰C and are not temperature sensitive compared to the Immortalized Human Mammary Fibroblasts (HMF3A) (abm, T0153).
Immortalization Method: Combined transduction with retroviruses carrying a temperature sensitive mutant (U19tsA58) of the SV40 Large T Antigen and a catalytic subunit of human telomerase (hTERT) BioSafety Level: II
Species: Human
Source Organ: Mammary Tissue
Growth Properties: Adherent
Morphology: Spindle shaped
Population Doubling: N/A
Seeding Density: Thaw entire contents into an appropriate T25 flask as specified in the Propagation instructions.
Markers: N/A
Donor Age: N/A
Donor Gender: N/A
Donor Ethnicity: N/A
Propagation: The base medium for this cell line is Prigrow III medium available at abm (TM003). To make the complete growth medium, add the following components to the base medium: fetal bovine serum (TM999)* to a final concentration of 10%, 1% L-glutamine (G275), and Penicillin/Streptomycin Solution (G255) to a final concentration of 1%. Carbon dioxide (CO2): 5%, Temperature: 37°C. The cells must be split 1:5 twice a week.* abm does not recommend to use heat-inactivated FBS for cell culture unless specified otherwise.
Quality Control: 1.) Telomerase-detection assay was used to detect telomerase activity. 2.) Flow cytometry confirmed expression of T antigen.
Shipping Condition: Dry Ice
Storage Condition: liquid nitrogen or -180°C