Fandom

VroniPlag Wiki

Pak/048

< Pak

31.340Seiten in
diesem Wiki
Seite hinzufügen
Diskussion0 Share

Störung durch Adblocker erkannt!


Wikia ist eine gebührenfreie Seite, die sich durch Werbung finanziert. Benutzer, die Adblocker einsetzen, haben eine modifizierte Ansicht der Seite.

Wikia ist nicht verfügbar, wenn du weitere Modifikationen in dem Adblocker-Programm gemacht hast. Wenn du sie entfernst, dann wird die Seite ohne Probleme geladen.

Functional characterization of the ‘PBX interacting protein’ (HPIP) in normal and malignant human haematopoiesis.

von Dr. Pak

vorherige Seite | zur Übersichtsseite | folgende Seite
Statistik und Sichtungsnachweis dieser Seite findet sich am Artikelende
[1.] Pak/Fragment 048 03 - Diskussion
Zuletzt bearbeitet: 2014-04-06 06:50:22 Hindemith
Ahmed 2007, Fragment, Gesichtet, Pak, SMWFragment, Schutzlevel sysop, Verschleierung

Typus
Verschleierung
Bearbeiter
SleepyHollow02
Gesichtet
Yes.png
Untersuchte Arbeit:
Seite: 48, Zeilen: 3-27
Quelle: Ahmed 2007
Seite(n): 40 f., Zeilen: 40: 18-34 - 41: 1-7
3.2.6. Purification of umbilical cord blood CD34+ cells (CB CD34+) from mononuclear umbilical cord blood (UCB).

Umbilical cord blood was collected in heparinised syringes according to institutional guidelines following normal full-term deliveries. Informed consent was obtained in all cases. Mononuclear cells (MNC) were separated using density gradient centrifugation. Fresh umbilical cord blood, not older than 12 hours, was diluted with 2 volumes of PBS and layered over Pancoll. Usually 35 ml of diluted blood was layered over 15 ml Pancoll in a 50 ml conical tube. This was centrifuged at 400x g for 30 minutes at 20°C in a swinging-bucket rotor without brakes. The upper layer was aspirated and discarded, leaving the interphase undisturbed. The interphase containing MNC such as lymphocytes, monocytes and thrombocytes was then transferred to a new 50 ml tube, washed twice with large volumes of PBS, and then counted before labelling with magnetic bead or fluorochrome conjugated antibodies.

hCB CD34+ cell purification was conducted using MACS CD34+ Cell Isolation Kit that uses positive selection method. Cells were resuspended in a volume of 300 ml per 1x108 MNCs These were blocked with 100 ml of FcR Blocking Reagent and labelled with 100 ml of CD34 Microbeads. When working with higher cell number, all the reagent volumes & the total volume was scaled up accordingly. This was followed by incubation for 30 minutes at 4-8°C. Cells were then washed twice by adding 10x the labelling volume of buffer and centrifuged at 300 x g for 15 minutes. The resultant cell pellet was then resuspended in 500 ml of MACS buffer and loaded into MS Column mounted on magnetic separator. The negative cells were allowed to pass through and the column was washed at least three times with 2 ml buffer.

3.2.6 Purification of human UCB CD133+ and CD34+ cells

Density Gradient Centrifugation:

Umbilical cord blood was collected in heparinised syringes according to institutional guidelines following normal full-term deliveries. Informed consent was obtained in all cases. Mononuclear cells (MNC) were separated using density gradient centrifugation. Fresh umbilical cord blood, not older than 12 hours, was diluted with 2 volumes of PBS and layered over Pancoll. Usually 35 ml of diluted blood was layered over 15 ml Pancoll in a 50 ml conical tube. This was centrifuged at 400x g for 30 minutes at 20°C in a swinging-bucket rotor without brakes. The upper layer was aspirated and discarded, leaving the interphase undisturbed. The interphase containing MNC such as lymphocytes, monocytes and thrombocytes was then transferred to a new 50 ml tube, washed twice with large volumes of PBS, and then counted before labelling with magnetic bead or fluorochrome conjugated antibodies.

Magnetic Separation:

CD133+ cell purification was conducted using MACS CD133 Cell Isolation Kit that uses positive selection method. Cells were resuspended in a volume of 300 μl per 108 cells, blocked with 100 μl of FcR Blocking Reagent and labelled with 100 μl of

[page 41]

Materials & Methods

CD133 Microbeads. When working with higher cell number, all the reagent volumes & the total volume was scaled up accordingly. This was followed by incubation for 30 minutes at 4-8°C. Cells were then washed twice by adding 10x the labelling volume of buffer and centrifuged at 300 x g for 15 minutes. The resultant cell pellet was then resuspended in 500 μl of MACS buffer and loaded into MS Column mounted on magnetic separator. The negative cells were allowed to pass through and the column was washed at least three times with 2 ml buffer.

Anmerkungen

The source is not given.

Sichter
(SleepyHollow02) Schumann


vorherige Seite | zur Übersichtsseite | folgende Seite
Letzte Bearbeitung dieser Seite: durch Benutzer:Hindemith, Zeitstempel: 20140406070908

Auch bei Fandom

Zufälliges Wiki