Browser Not Supported.

IE users: please get the latest browser version at Microsoft Internet Explorer Home Page

if yot already have the latest browser version please verify that your browser settings is not in compatibility mode

Browser: unknown
Version: 0

All in Vitro Cells > Adult Stem / Progenitor Cell card

Peripheral blood-derived hematopoietic stem cells (Imperial College London)

Granulocyte colony-stimulating factor (GCSF)-mobilized peripheral blood cells were obtained from excess leukapheresis samples processed by the Stem Cell Laboratory, Hammersmith Hospital. Informed consent and local research ethics committee approval were granted in all cases.

CD34 cells were diluted 1:4 in Hanks’ buffered saline solution (HBSS) before mononuclear cells (MNCs) were separated, by centrifugation, over a Lymphoprep density gradient at 1,800 rpm for 30 minutes. The MNC fraction was collected and first washed in HBSS, then in MACS (magnetic cell sorting) buffer (phosphate-buffered saline buffer supplemented with bovine serum albumin (0.5%) and EDTA (5 mM, pH 7.2)). CD34 cells were isolated from MNCs, using the CD34-positive cell selection kit. Isolated CD34 cells were plated on 35-mm2 Petri dishes in α -minimal essential medium (α-MEM) supplemented with fetal bovine serum (FBS; 15%) and incubated for 2 hours at 37°C in 5% CO2. After 2 hours, the nonadherent cell fraction was removed by washing the plates three times. Adherent CD34 cells were cultured in α-MEM supplemented with FBS (30%) and cytokines (20 ng/ml stem cell factor [SCF], GM-SCF (1ng/ml), IL-3 (5 ng/ml), and G-CSF (100 ng/ml)) at 37°C in 5% CO2 in air.

See additional Stem, Progenitor & Primary Cells for: Blood
Adult Stem / Progenitor Cell
Homo sapiens
Peripheral blood-derived hematopoietic stem cells