Immortalized Human Mammary Fibroblasts (HMF3A)

Immortalized Human Mammary Fibroblasts (HMF3A) were generated through the combined transduction of mammary fibroblasts with a temperature sensitive mutant (U19tsA58) of the SV40 Large T Antigen and a catalytic subunit of human telomerase (hTERT). The HMF3A cells grow best at a temperature of +33.5⁰C and undergo complete arrest at +39.5⁰C. These cells are temperature sensitive unlike the Immortalized Human Mammary Fibroblasts (HMF3S) (abm, T0154). These conditionally immortalized cells are suitable for cell engineering and are theoretically capable of generating immortal diploid human lines.

Immortalization Method: Combined transduction with retrovirus 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: +33.5°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