If Using Stable Producer Cell Line

1. Producer cell line for generation of ecotropic retrovirus encoding lacZ: CRE BAG 2 (American Type Culture Collection, Accession Number CRL 1858).

2. Growth media (GM, store at 4°C): The following are added to DMEM (formulation containing 4500 mg/L glucose and 2 mM L-glutamine) to give the final indicated concentrations: 10% fetal bovine serum (FBS); 100 U/mL penicillin; 1000 ||g/mL streptomycin.

3. Passaging cells: Phosphate-buffered saline-calcium and magnesium free (PBS-CMF), pH 7.4, Trypsin (0.05% trypsin/0.53 mM EDTA).

4. Freezing media: 90% calf serum (see Note 1) and 10% dimethylsulfoxide (DMSO).

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