Single nucleated cells (MNCs) acquisition assay

Summary

Source: Human Stem Cell Culture

Operation method

basic program

Materials and Instruments

Cord blood
Sterilized appropriate culture medium or freezing solution PBSA
Ficoll-Hypague (density 1.077 g L) pasteurized pipette 50 mL centrifuge tube

Move

(a) Cord blood samples were diluted 1:1 with PBSA.

(b) Inhale 15mL Ficoll-Hypague into a 50mL centrifuge tube.

(c) Slowly add 30mL PBSA-diluted cord blood sample to Ficoll-Hypague.

(d) Centrifuge at 450g at room temperature for 20~30min.

(e) After centrifugation, a layer of cells, a layer of single nucleated cells with a lower density than that of the FicoIlHypague fluid, was clearly visible on the surface of the Ficoll-Hypague fluid.

(f) Using a pasteurized pipette, the single nucleated cell layer was aspirated and transferred to another centrifuge tube.

(g) Wash the cells with PBSA and centrifuge at 200g for 10min at room temperature.

(h) Pour off the supernatant, resuspend the cells with PBSA, wash the cells and repeat the above steps (g).

(i) Finally, resuspend the cells with suitable medium (chilled liquid or medium)

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