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Immortalized-Human-Mammary-Fibroblasts-(HMF3A)

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Immortalized-Human-Mammary-Fibroblasts-(HMF3A)

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Immortalized-Human-Mammary-Fibroblasts-(HMF3A)

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	Print Version
BioSafety Level	II
Organism	Homo sapiens
Source Organ	Mammary Tissue
Growth Properties	Adherent
Morphology	Spindle
Recommended Seeding Density	Thaw entire contents into an appropriate T25 flask as specified in the Propagation instructions.
Applications	For Research Use Only
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)
Description	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 33.5?C and undergo complete arrest at 36.5?C. 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.
Procedure Overview
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Propagation	Use of PriCoat? 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 with the following conditions. The base medium for this cell line is Prigrow III medium available from 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% glutamate, and Penicillin/Streptomycin(G255). Atmosphere: air: 95%, CO2: 5%; Temperature: 33.0°C. The cells must be split 1:5 twice a week.
Quality Control	1.) Telomerase-detection assay was used to detect telomerase activity. 2.) Flow cytometry confirmed expression of T antigen.
Disclaimer	1. The CoA for this product (provided upon request) verifies the cell-type specific gene expression via RT-PCR only. 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 μg, Cat.# C207, $450.00) or cell lysate (100 μg, Cat.# C206, $600.00) to perform preliminary experiments, or alternatively use our Gene Expression Assay Service (Cat# C138). All sales are final. 2. We strongly 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). 3. 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. 4. 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.