Materials and Reagents

1. 27-gauge needle (TERUMO, catalog number: NN-2719S )
   
2. 1 ml syringe (TERUMO, catalog number: SS-01T )
   
3. 6-well plate (Coring, Falcon^®, catalog number: 353046 )
   
4. 50 ml tube
   
5. 70 μm cell strainer (Coring, Falcon^®, catalog number: 352350 )
   
6. 10 ml syringe (TERUMO, catalog number: SS-10SZ )
   
7. Mice (adult > 6 weeks, any sex, any strain)
   
8. Hyaluronidase (60 mg/ml stock) (Sigma-Aldrich, catalog number: H3506 )
   
9. Collagenase type VIII (10 mg/ml stock) (Sigma-Aldrich, catalog number: C2139 )
   
10. RPMI 1640 (Sigma-Aldrich, catalog number: R8758 )
    
11. Fetal bovine serum, heat inactivated (Biowest, catalog number: S1820-500 )
    
12. HANK’s solution “Nissui”② (NISSUI PHARMACEUTICAL, catalog number: 05906 )
    
13. NaHCO₃ (NAKARAI TESQUE, catalog number: 31212-25 )
    
14. NaN₃ (NAKARAI TESQUE, catalog number: 31208-82 )
    
15. Digestion medium (see Recipes)
    
16. FACS solution (see Recipes) 
    

Equipment

1. Surgical scissors and tweezers
   
2. Mini-shaker 3D (Biosan, catalog number: BS-010151-AAK )
   
3. Refrigerated centrifuge
   

Procedure