abm | Immortalized Mouse Microglia (SIM-A9) | T0247

abm | Immortalized Mouse Microglia (SIM-A9) | T0247

Immortalization Method:

Spontaneous immortalization

BioSafety Level:

II

Organism:

C57BL/6 mouse

Species:

Mouse

Source Organ:

Postnatal mouse cerebral cortices

Organ Type:

Brain

Growth Properties:

Adherent

Morphology:

Round or Polygonal

Passage Number:

N/A

Population Doubling:

30 - 40 hours

Seeding Density:

45,000 - 50,000 cells/cm10²; Recommended split ratio is 1:2

Markers:

CD68, Iba1

Donor Age:

N/A

Donor Gender:

N/A

Donor Ethnicity:

N/A

Propagation:

The base medium for this cell line is Prigrow IV medium available at abm . To make the complete growth medium, add the following components to the base medium: heat-inactivated fetal bovine serum to a final concentration of 10% donor horse serum (Thermo Fisher Scientific; Cat. no. 1605013) to a final concentratio of 5% and Penicillin/Streptomycin Solution to a final concentration of 1%.

Quality Control:

1) Functional assays such as the nitrate assay was performed to assess nitric oxide production; 2) ELISA was used to determine the TNFα secretion after inflammatory stimulation; 3) Western blot and immunocytochemistry were used to confirm the expression of microglial specific markers such as CD68 and Iba1 and M1/M2 phenotype-associated protein expression (COX-2/iNOS and Arg-1) after stimulation; 4) Aβ_1-42 or E. coli-derived bioparticles uptake was used to determine phagocytic activity of SIM-A9 cells.

Shipping Condition:

Dry Ice

Storage Condition:

liquid nitrogen or -180C