Immortalized Mouse Embryo Neural Stem Cells (MuNSC)

Cells were derived from 13.5 dpc murine embryo brain and immortalized by the 3T3 protocol. Neurospheres can be induced from monolayer cells by growing them in Prigrow III medium available at abm, Cat. No. TM003, supplemented with B27 supplement (Gibco) to a final concentration of 1%, epidermal growth factor (Z200025) to a final concentration of 10 ng/mL, basic fibroblast growth factor-2 (Z200015) to a final concentration of 10 ng/mL, 100U/mL penicillin, and 100 ug/mL streptomycin. Carbon dioxide (CO₂): 5%, Temperature: +37.0°C. Monolayer cells expressing specific neural lineage markers can be differentiated from neurospheres when grown in regular Prigrow III propagation media. Carbon dioxide (CO₂): 5%, Temperature: +37.0°C. Neural stem cells can be used to study neurodegenerative diseases and research neural repair mechanisms.

Immortalization Method: Immortalized by using the 3T3 protocol.

BioSafety Level: II

Organism: Mouse (M. musculus)

Species: Mouse

Source Organs: Brain

Organ Type: Brain

Growth Properties: Adherent

Morphology: Fibroblast-like

Markers: GFAP, NESTIN, and MBP

Donor Age: 13.5 dpc embryo

Propagation: The base medium for this cell line is Prigrow III medium available at abm, Cat. No. TM003. To make the complete growth medium, add the following components to the base medium: fetal bovine serum (TM999)* to a final concentration of 10%, L-glutamine (G275) to a final concentration of 2 mM and Penicillin/Streptomycin Solution (G255) to a final concentration of 1%.Change media every 3-4 days.Carbon dioxide (CO₂): 5%, Temperature: +37.0°C.* Do not use heat-inactivated FBS for cell culture unless specified otherwise.

Quality Control: 1) Senescenceβ-galactosidase staining kit; 2) Western blot; 3) Immunocytochemistry; 4) Cell proliferation assays; 5) Wound-healing assay; 6) Metaphase chromosome preparation

Shipping Condition: Dry Ice

Storage Condition: liquid nitrogen or -180°C

Reference: Almengló, C., González-Mosquera, T., Caamaño, P., Seoane, M., Fraga, M., Devesa, J., Costoya, J. A., & Arce, V. M. (2020). Immortalization of a cell line with neural stem cell characteristics derived from mouse embryo brain. Developmental dynamics : an official publication of the American Association of Anatomists, 249(1), 112–124. https://doi.org/10.1002/dvdy.103