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$Immortalized-Human-Adipose-derived-Stromal-Cells---Bmi-1\/hTERT$

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Immortalized-Human-Adipose-derived-Stromal-Cells---Bmi-1\/hTERT

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Immortalized-Human-Adipose-derived-Stromal-Cells---Bmi-1\/hTERT

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	Print Version
BioSafety Level	II
Organism	Homo sapiens
Source Organ	Adipose Tissue
Donor Age	30
Donor Gender	Female
Growth Properties	Adherent
Population Doubling	Expected doubling time of 4 days
Recommended Seeding Density	6,000-8,000 cells/cm^2
Markers	CD44, CD73, CD90 and CD105
Applications	For Research Use Only
Immortalization Method	Serial passaging and transduction with recombinant lentiviruses carrying murine Bmi-1 [Ref. seq.: NM_007552.4] and hTERT [Ref.seq.: NM_198253.2] gene
Description	With its ability to maintain multipotency, mesenchymal stem cells provide new venues for cell-based therapy in restoration of damaged or diseased tissue. Most clinical studies utilize bone marrow-derived stem cells, however adipose-derived stromal cells (ASCs) is emerging as an alternative source for their ease of harvesting. The Immortalized Adipose-derived Stromal Cells- Bmi-1/hTERT is derived from human ASCs that have maintained their primary phenotypes and their ability to respond to osteogenic and adipogenic inductions. After 3 weeks of induced osteogenic differentiation, the immortalized ASCs show nuclear translocation of RUNX2, upregulation of alkaline phosphatase and calcium deposition. Furthermore, the immortalized ASCs-Bmi-1/hTERT exhibit normal karyotype and has been shown to exceed 140 population doublings, making these cells suitable for studies in regenerative medicine.
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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 IV medium available from ABM (TM004). To make the complete growth medium, add the following components to the base medium: fetal bovine serum (TM999) to a final concentration of 10%, L-glutamine and antibiotics (gentamicin). Optional for better growth: 1-5 ng/mL human recombinant bFGF .Atmosphere: air: 95%, CO₂: 5%; Temperature: 37.0°C
Quality Control	1. Real Time PCR, immunofluorescence, and telemorase activity was used to quantify Bmi-1 and/or hTERT gene expression in the immortalized cell line. 2. Culture tested negative for mycoplasma
Reference	Nemet, K., et al (2012). Combined introduction of Bmi-1 and hTERT immortalizes human adipose tissue-derived stromal cells with low risk of transformation. Biochemical and Biophysical Research Communications (2012) 422, 28-35.
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.