Description: SIRMu-1 is an immortalized rat Müller-1 cell line, monoclonal, derived from 2 sequential single cell/clonal isolations.
Note: Müller cells are a major type of glial cells in the retina of the eye.

Organism: Rat (Rattus norvegicus)

Strain: Sprague-Dawley

Tissue: Retina

Growth Properties: Adherent

Morphology: Epithelial-like

Growth Medium: EMEM + 20% Foetal Calf Serum (FCS) + 25mM Glucose + 2mM L-Glutamine

Resuscitation: Remove protective cryoflex layer around the ampoule prior to thawing. Thaw the ampoule by gently agitating in a 37°C waterbath; thawing should be rapid (around 2 minutes). A centrifugation step to remove the cryoprotectant after thawing is necessary for this cell line.

NOTE: This cell line may take up to 48hours to seed.

Subculturing Procedure:

Medium Renewal: 2-3 times per week

Subcultivation Ratio:  1:5 – 1:10 split subconfluent cultures (70-80%). Harvest the cells using 0.05% Trypsin/EDTA at 37°C for 5 minutes.

At thaw, the cells should be seeded at a density of 2-3 x 10⁴cells/cm². After recovery from thaw (48-72 hours), they can be seeded at 0.5-1.0 x 10⁴cells/cm².

Flasks will require passaging twice weekly at a 1:10 seeding density.

Culture Conditions: Incubate the culture at 37°C with 5% CO₂.

Cryoprotectant Medium: 10% DMSO + 90% FCS.

Handling Procedure for Frozen Cells: Upon receipt, frozen ampoules should be transferred directly to liquid nitrogen storage without delay, if not to be used immediately. Storage at -80°C may result in loss of viability.

Additional Information: 

The cells express the Müller and/or glial cells markers:

• Glutamine Synthetase
• S-100 Protein
• Vimentin
• Glial Fibrillary Acidic Protein (GFAP)

Gender: Male

Depositor: Dan Peet – University of Adelaide, South Australia

References:

Kittipassorn T, Haydinger CD, Wood JPM, Mammone T, Casson RJ, Peet DJ.

Characterisation of the novel spontaneously immortalised rat Müller cell line SIRMu-1.

Exp Eye Res. 2019 Apr; 181:127-135

PubMed: 30677389

Cellosaurus: CVCL_UW38