Isolation of Human Blood Progenitor and Stem Cells from Peripheral Blood by Magnetic Bead

Materials and Reagents

1. CD34⁺ cells
   
2. Peripheral blood sample (at least 50 ml)
   
3. Dextran solution (Sigma-Aldrich, catalog number: D1037-500G )
   
4. PBS (Life Technologies, Invitrogen™, catalog number: 14190-094 )
   
5. Ficoll human (PAA Laboratories GmbH, catalog number: P04-60500 )
   
6. Fetal calf serum (FCS) (Hyclone, catalog number: SV30160.03 )
   
7. CD34 MicroBeads and FcR blocking reagent (Miltenyi Biotec, catalog number: 130-046-702 )
   
8. APC mouse anti-human CD34 antibody (BD Biosciences, catalog number: 555824 )
   
9. EDTA
   
10. 2% dextran solution
    
11. 1 L 2% dextran solution (see Recipes)
    

Equipment

1. Centrifuges
   
2. Auto MACS Pro separator (Miltenyi Biotec)
   
3. 30 μm nylon mesh (Miltenyi Biotec, catalog number: 130-041-407 )
   
4. Tissue culture hood
   
5. Flow cytometer
   
6. Vacuum filter unit (22 μm, GP Millipore Express PLUS membrane)
   

Procedure