|
| Status |
Public on Dec 10, 2008 |
| Title |
Monoblasts_1 |
| Sample type |
RNA |
| |
|
| Source name |
In vitro differentiated normal human monoblasts
|
| Organism |
Homo sapiens |
| Characteristics |
type: in vitro differentiated cells derived from CD34+ stem/progenitor cells from healthy donors
|
| Growth protocol |
Monoblasts (CD14+ precursors) and myeloblasts (CD14- precursors) were obtained by in vitro differentiation of cord blood (CB) derived CD34+ cells performed as already described by Montanari et al. Briefly, CB CD34+ cells were cultured in IMDM added with 20% FCS (Bio-Whittaker, Walkersville, MD, USA), in the presence of human hematopoietic cytokines: SCF (50 ng/ml), Flt3-ligand (Flt3-l) (50 ng/ml), IL-11 (50 ng/ml), IL-6 (10 ng/ml), IL-3 (10 ng/ml) and G-CSF (10 ng/ml) (all from R&D Systems, Minneapolis, MN, USA). After 7 days of culture, hematopoietic cells were analyzed, by flow cytometry, for CD14 antigen expression, estimated at about 25-30% of the entire cell population. Then monoblasts (CD14+) and myeloblasts (CD14-) cell fractions were obtained by immunomagnetic separation using the MACS technology (Miltenyi). Differentiation of CD34+ cells was monitored by morphological analysis of MGG-stained cytospins and by flow-cytometric analysis of CD34, CD38 and CD14 surface antigen expression.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total cellular RNA was isolated by means of RNeasy Mini Kit (Qiagen, Valencia, CA) following manufacturer's recommendations.
|
| Label |
biotin
|
| Label protocol |
RNA samples from cells of different donors were pooled to obtain 5 μg of total cellular RNA that were used for target synthesis according to the standard one-cycle Affymetrix protocol, supplied by the manufacturer (Affymetrix, Santa Clara, CA).
|
| |
|
| Hybridization protocol |
Following fragmentation, 15 ug of cRNA were hybridized for 16 hr at 45C on GeneChip HG-U133A Array. GeneChips were washed and stained in the Affymetrix Fluidics Station 450.
|
| Scan protocol |
GeneChips were scanned using the Affymetrix GeneChip Scanner GCS3000.
|
| Description |
In vitro differentiated normal human monoblasts
|
| Data processing |
The data were analyzed using Bioconductor packages and RMA pre-processing procedure
|
| |
|
| Submission date |
Sep 16, 2008 |
| Last update date |
Sep 17, 2008 |
| Contact name |
Francesco Ferrari |
| E-mail(s) |
ferrari.francesco@unimore.it
|
| Phone |
+39(0)592055512
|
| Fax |
+39(0)592055410
|
| URL |
http://www.xlab.unimo.it
|
| Organization name |
University of Modena and Reggio Emilia
|
| Department |
Department of Biomedical Sciences
|
| Lab |
Experimental genomics & transcriptomics laboratory
|
| Street address |
Via G. Campi 287
|
| City |
Modena |
| ZIP/Postal code |
41100 |
| Country |
Italy |
| |
|
| Platform ID |
GPL96 |
| Series (2) |
| GSE12803 |
Gene expression in human myeloid cells |
| GSE12837 |
Gene expression in human myeloid cells. |
|
