Immortalized Mouse Conditionally Reprogrammed Otic Stem Cells (CR-OSC)

Otic Stem Cells are auditory progenitor cells which can be differentiated into cells which resemble auditory hair cells (HC) and express HC genes. Immortalized Mouse Conditionally Reprogrammed Otic Stem Cells (CR-OSC) were immortalized through exposure to conditional reprogramming conditions and their growth rate remained consistent for 10 passages. The cells upregulate HC genes in response to differentiation conditions, regardless of passage number. The Immortalized Mouse Conditionally Reprogrammed Otic Stem Cells (CR-OSC) are useful in the field of audiology. The applications of this cell line include monitoring and manipulating HC specific genes and experiments involving genetic mutations, knock-outs or overexpression, both of which are enhanced by the large number of cells this cell line produces.

Immortalization Method: Conditional reprogramming

BioSafety Level: II

Species: Mouse

Source Organ: Organ of Corti

Growth Properties: Adherent

Morphology: Polygonal

Population Doubling: N/A

Seeding Density: N/A

Markers: prestin, Atoh1, Pou4f3, myosin VI, myosin VIIa, parvalbumin, otoferlin, and VGlut3

Donor Age: N/A

Donor Gender: N/A

Donor Ethnicity: N/A

Propagation: Grow cells in 6-well plate coated with mitomycin c (Enzo Life Sciences) inactivated (10 ug/mL, 3 hours) 3T3-J2 cells (6 x 10⁵ cells/well) with the following conditions. The base medium for this cell line is Prigrow III medium available at abm (TM003). To make the completed growth medium, add the following components to the base medium: fetal bovine serum (TM999)* to final concentration of 5%, 0.4 ug/mL hydrocortisone (Sigma-Aldrich), 5 ug/mL Insulin Solution (TM053), 8.4 ng/mL cholera toxin (Sigma-Aldrich), 10 ng/mL epidermal growth factor (Invitrogen), 24 ug/mL adenine (Sigma-Aldrich), and 5 uM Y-27632 (ROCK inhibitor, Enzo Life Sciences).
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) Immunohistochemistry to detect upregulation of genes in response to differentiation conditions; 2) Viral transduction of Atoh1 overexpression to detect gene expression response; 3) Thyroid hormone treatment to detect gene expression response

Shipping Condition: Dry Ice

Storage Condition: liquid nitrogen or -180°C