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

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.

Immortalization Method: Serial passaging and transduction with recombinant lentiviruses carrying mouse Bmi-1 (pLOX-CWBmi1) [Ref. seq.: NM_007552.4] and hTERT (pLOX-TERT-iresTK) [Ref.seq.: NM_198253.2] gene

BioSafety Level: II

Species: Human

Source Organ: Adipose Tissue

Growth Properties: Adherent

Morphology: N/A

Population Doubling: 90-100 Hours

Seeding Density: 6,000-8,000 cells/cm²

Markers: CD44, CD73, CD90 and CD105

Donor Age: 30

Donor Gender: Female

Donor Ethnicity: N/A

Propagation: The base medium for this cell line is Prigrow IV medium available at 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%, 2mM L-glutamine (G275) and 50 ug/ml gentamicin (G272). Optional for better growth: 1-5 ng/mL human recombinant bFGF (Z101455).
Carbon dioxide (CO₂): 5%, Temperature: +37.0°C. * abm does not recommend to use heat-inactivated FBS for cell culture unless specified otherwise.

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

Shipping Condition: Dry Ice

Storage Condition: liquid nitrogen or -180°C