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Typus
Verschleierung
Bearbeiter
SleepyHollow02
Gesichtet
Yes.png
Untersuchte Arbeit:
Seite: 49, Zeilen: 1-8
Quelle: Ahmed 2007
Seite(n): 41, Zeilen: 7-19
[The column was then removed from the separator, placed on a collection tube, loaded with fresh buffer, and the magnetically labelled cells flushed out using the] plunger. The magnetic separation was usually repeated to get a purity of more than 95%. Purified cells were then frozen in FBS with 10% DMSO and thawed when needed for pre-stimulation and transduction. hCB CD34+ cell enrichment was done by FACS. For separation by FACS, MNCs were thawed from frozen stocks or prepared freshly from UCB and labelled using anti CD34-PE antibody (100 ml per 108 cells), for 30 minutes on ice. Labelled cells were then washed twice with PBS, resuspended in FACS buffer and sorted. The sorted cells with purity above 95 % were used for 48 hour pre-stimulation followed by transduction. The column was then removed from the separator, placed on a collection tube, loaded with fresh buffer, and the magnetically labelled cells flushed out using the plunger. The magnetic separation was usually repeated to get a purity of more than 95%. Purified cells were then frozen in FBS with 10% DMSO and thawed when needed for pre-stimulation and transduction.

FACS sorting:

CD34+ cell enrichment was done either by MACS as done for CD133+ cells or by FACS. For separation by FACS, MNCs were thawed from frozen stocks or prepared freshly from UCB and labelled using anti CD34-PE antibody (100 μL per 108 cells), for 30 minutes on ice. Labelled cells were then washed twice with PBS, resuspended in FACS buffer and sorted. The sorted cells with purity above 95% were used for 48 hour pre-stimulation followed by transduction.

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(SleepyHollow02) Schumann

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