Immortalized Mouse Conditionally Reprogrammed Otic Stem Cells (CR-OSC) | T0372

Immortalized Mouse Conditionally Reprogrammed Otic Stem Cells (CR-OSC) | T0372

Immortalization Method:

Conditional reprogramming

BioSafety Level:

II

Organism:

Mus musculus

Species:

Mouse

Source Organ:

Organ of Corti

Organ Type:

Auditory

Growth Properties:

Adherent

Morphology:

Polygonal

Passage Number:

N/A

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 . To make the completed growth medium, add the following components to the base medium: fetal bovine serum 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 -180C