Immortalized Human Brown Pre-adipocytes (PAZ6)

Immortalized Human Brown Pre-Adipocytes are derived from infant brown adipose tissue. The vascular stromal cells were immortalized through microinjection of the vector pUC 18 carrying SV40 large and small T antigen under the control of a truncated human vimentin promoter. These immortalized cells express both PPAR? and UPC1 and can be differentiated using a drug cocktail described below, resulting in terminally differentiaded brown adipocytes that display similar expression profiles and morphology to primary brown adipocytes. The functionality of this line may make it useful as an in vitro model of brown adipocytes and preadipocytes study

Immortalization Method: SV40 large and small T-antigen

BioSafety Level: II

Species: Human

Source Organ: Adipose Tissue

Growth Properties: Adherent

Morphology: Epithelial

Population Doubling: N/A

Seeding Density: N/A

Markers: N/A

Donor Age: Infant

Donor Gender: N/A

Donor Ethnicity: N/A

Propagation: The base medium for this cell line is Prigrow IV medium available from abm, Cat. No. TM004. To make the completed growth medium, add the following components to the base medium: fetal bovine serum (TM999)* to a final concentration of 8% , 1% Glutamax and 1% Penicillin/Streptomycin Solution (G255).
Carbon dioxide (CO₂): 5%, Temperature: +37.0°C.
For differentiation of pre-adipocytes to adipocytes, the following media composition is recommended:
A base medium of Prigrow IV media from abm (TM004) with fetal calf serum to a final concentration of 5%, 15 mM of HEPES, 33µM Biotin, 17µM Pantothenate, 1nM Triiodothyronine, 100nM Dexamethoasone, 1µM Pioglitazone, 500 nM Insulin, 0.25mM IBMX ( for the first 4 days of differentiation) and 1% Penicillin/Streptomycin (G255).
Differentiation of cells takes approximately 14 days. During this time, the cells should be washed with PBS and the differentiation media change every 2 days.
Adipocytes do not freeze well. Thus it is suggested that these cells initially be expanded and back up vials produced for recovery purposes.
* abm does not recommend to use heat-inactivated FBS for cell culture unless specified otherwise.

Quality Control: 1) Compared preadipocyte and adipocyte expression after terminal differentiation via semi-quantitative RT-PCR (Figure 1). 2) Visualized morphology of differentiated and undifferentiated cells via phase contrast microscopy (Figure 2). 3) Compared the expression profiles of primary and differentiated t0729 cells (Figure 3).

Shipping Condition: Dry Ice

Storage Condition: liquid nitrogen or -180°C