Immortalized Mouse Subcutaneous Adipose Multipotent Mesenchymal Cells (m17.ASC-GFP)

Immortalized Mouse Subcutaneous Adipose Multipotent Mesenchymal Cells (m17.ASC-GFP) were genetically modified by lentiviral vectors carrying green fluorescent protein
(GFP). When injected in the spleen of NOD/SCID-gnull monocrotaline-treated mice, m17.ASC-GFP cells efficiently engrafted in the liver, candidating this cell line as a valuable stem cell model.
It stably expresses the stem cell markers Sca-1, c-Kit/CD117, nestin, nucleostemin, CD44, and CD106, but is negative for the embryo stem cell markers Nanog, Oct4, and Sox2. The cells were endowed with multipotency and can be induced to differentiate toward osteogenic, chondrogenic, adipogenic, and cardiogenic phenotypes. It displays a normal karyotype and stable telomeres, neither proliferates when plated in soft agar nor gives rise to tumors when injected subcutaneously in NOD/SCID-g null mice. The cell line is thus recommended as a valuable tool for a number of applications in ex vivo tissue engineering to study the differentiation mechanisms involved in tissue repair, as well as a model for both pharmacological and toxicological studies.

abm also offers the Immortalized Mouse Subcutaneous Adipose Multipotent Mesenchymal Cells (m17.ASC) (abm, T0542) which does not carry GFP expression.

Immortalization Method: Spontaneous immortalization

BioSafety Level: II

Species: Mouse

Source Organ: Adipose tissue

Growth Properties: Adherent

Morphology: Fibroblast-like

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 the cell line is Claycomb medium (Sigma-Aldrich). To make the completed growth medium, add the following components to the base medium: 2mM L-glutamine (G275), fetal bovine serum (TM999)* to a final concentration of 10%, Penicillin/Streptomycin Solution (G255) to a final concentration of 1%.
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) RT-PCR and Cytofluorimetry were performed on the cDNA from the cells to confirm the expression of stem cell markers Sca-1, c-kit, Islet 1, nestin, and nucleostemin.
2) Cells after differentiation were specifically stained with alizarin red, adipo-red and Alcian blue, and the tissue-specific collagen II antibodies, respectively to confirm osteogenic, adipogenic, and chondrogenic phenotypes the cells’ multipotency.
3) G-banding analysis for macroscopic chromosome alterations over time compared with normal mouse karyotypes.
4) GFP transduction was confirmed by culturing with antibiotics.

Shipping Condition: Dry Ice

Storage Condition: liquid nitrogen or -180°C