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Sample GSM25884 Query DataSets for GSM25884
Status Public on Apr 06, 2005
Title lin+CD34+ II replicate
Sample type RNA
 
Source name lin+CD34+ progenitor cells
Organism Homo sapiens
Extracted molecule total RNA
 
Description CD34+ lineage positive progenitors cells purified from peripheral blood.
To obtain PB progenitor cells, 12 healthy donors received glycosylated recombinant human (rh) G-CSF (Lenograstim, Rhone-Poulenc Rorer, Milan, Italy) administered subcutaneously at 10 g/kg/d for 5 6 d. Leukaphereses were performed on days 5 and 6 as previously described (Lemoli et al, 1997; Lemoli et al, 2000). Haematopoietic progenitor purification and subsequent studies were always performed using day-5 collections at the peak time of PB CD34+ cells. The protocol was approved by the ethical committee of the University Hospital and each donor gave written informed consent.
Mononuclear light density (<1·077 g/ml) cells (MNC) were enriched using Ficoll-Paque (Pharmacia, Uppsala, Sweden) and resuspended in 1% bovine serum albumin (BSA; Sigma Chemical Co, St Louis, MO, USA).
CD34+ cells were highly purified by MiniMacs high-gradient magnetic separation column (Miltenyi Biotec, Bergisch Gladbach, Germany) according to the manufacturer's instructions. To assess the percentage of CD34+ elements, aliquots of the CD34+ target cells were restained with an antibody (HPCA-2; IgG1a-fluorescein isothiocyanate [FITC]; Becton Dickinson, San Jose, CA) directed at a different epitope of CD34 antigen than that (QBend10) used with the MiniMacs system. Briefly, CD34+ cells were incubated for 30 minutes in the dark at 4°C with HPCA-2-FITC. Propidium iodide (2 µg/mL) was added for the detection of nonviable cells, which were excluded from analysis. After two washes in PBS/BSA, flow cytometric analysis was performed on a gated population set on scatter properties by using FACScan equipment (Becton Dickinson). A minimum of 10,000 events was collected in list mode on FACScan software. (Lemoli et al, 1997). The percentages of CD34+ cells in PB samples were 1·6±1%, and 0·9±0·3% of the MNC fraction respectively. After magnetic separation, the percentages of CD34+ in PB samples were 95±5%.

Keywords = lin+CD34+
Keywords = hemopoietic progenitors cells
 
Submission date Jun 21, 2004
Last update date Mar 16, 2009
Contact name Rossella Manfredini
E-mail(s) manfredini.rossella@unimore.it
Phone +390592058065
Organization name Centre for Regenerative Medicine
Department Life Sciences
Street address Via Gottardi 100
City Modena
ZIP/Postal code 41100
Country Italy
 
Platform ID GPL8300
Series (1)
GSE1493 Hematopoietic stem cell subsets

Data table header descriptions
ID_REF
VALUE Signal CD34+lin+ II
ABS_CALL Detection CD34+lin+ II

Data table
ID_REF VALUE ABS_CALL
AFFX-YEL024w/RIP1_at 0.8 A
AFFX-YEL021w/URA3_at 0.4 A
AFFX-YEL018w/_at 0.5 A
AFFX-YEL002c/WBP1_at 8.5 A
AFFX-TrpnX-M_at 0.8 A
AFFX-TrpnX-5_at 7.6 A
AFFX-TrpnX-3_at 0.4 A
AFFX-ThrX-M_at 0.5 A
AFFX-ThrX-5_at 0.7 A
AFFX-ThrX-3_at 1.8 A
AFFX-PheX-M_at 1 A
AFFX-PheX-5_at 0.4 A
AFFX-PheX-3_at 5.5 A
AFFX-MurIL4_at 1 A
AFFX-MurIL2_at 2.5 A
AFFX-MurIL10_at 8.1 A
AFFX-MurFAS_at 4.3 A
AFFX-M27830_M_at 56.6 A
AFFX-M27830_5_at 63 P
AFFX-M27830_3_at 51 A

Total number of rows: 12625

Table truncated, full table size 197 Kbytes.




Supplementary data files not provided

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