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Series GSE40790 Query DataSets for GSE40790
Status Public on Apr 26, 2013
Title Transgene-free iPSCs generated from small volume peripheral blood nonmobilized CD34+ cells
Organism Homo sapiens
Experiment type Methylation profiling by genome tiling array
Summary A variety of somatic cells can be reprogrammed to induced pluripotent stem cells (iPSCs), but the small number of CD34+ hematopoietic stem cells (HSCs) present in non-mobilized peripheral blood (PB) would be a convenient and desirable starting target. We report here a simple method for targeting derivation of iPSC from non-mobilized PB CD34+ HSCs using immunobead purification and 2-4 day culture to achieve enrichment of CD34+ HSCs to 80±9%, followed by reprogramming transduction with loxP-flanked polycistronic (Oct4, Klf4, Sox2, and c-Myc) STEMCCA-loxP lentivector at an MOI of 2. Our yield was 4.7±2.2 iPSC colonies (n=12) per 20 mL non-mobilized peripheral blood, where most colonies had single copy STEMCCA-loxP that was easily excised by transient transfection expression of Cre. Resultant iPSC clones expressed pluripotent cell markers, had genomic methylation pattern closely matching embryonic stem cells, and generated teratomas containing tissues of all three germ lineages in immunodeficient mice. Furthermore, we conclude that these iPSC are derived from the non-mobilized CD34+ HSCs enriched from PB rather than from any lymphocyte or monocyte contaminants because they lacked somatic rearrangements typical of T or B lymphocytes, and because we demonstrated that purified CD14+ monocytes do not yield iPSC colonies under these reprogramming conditions.
Overall design Comparison of pluripipotent, induced pluripotent, and differentiated cell types
Contributor(s) Merling RK, Sweeney CL, Choi U, DeRavin SS, Myers TG, Otaizo-Carrasquero F, Pan J, Linton G, Chen L, Koontz S, Theobald NL, Malech HL
Citation(s) 23386128
Submission date Sep 11, 2012
Last update date Mar 22, 2019
Contact name Timothy G Myers
Organization name National Institute of Allergy and Infectious Diseases
Department Research Technologies Branch
Lab Genomic Technologies Section
Street address 50 South Drive, Room 5509
City Bethesda
State/province MD
ZIP/Postal code 20892-8005
Country USA
Platforms (1)
GPL13534 Illumina HumanMethylation450 BeadChip (HumanMethylation450_15017482)
Samples (21)
GSM1139402 A6122 iPSC / iGP91-02
GSM1139403 A6123 iPSC / iNC-01
GSM1139404 A6126 ESC / H9
BioProject PRJNA174964

Download family Format
SOFT formatted family file(s) SOFTHelp
MINiML formatted family file(s) MINiMLHelp
Series Matrix File(s) TXTHelp

Supplementary file Size Download File type/resource
GSE40790_RAW.tar 183.1 Mb (http)(custom) TAR
GSE40790_unmethylated_methylated_intensities.txt.gz 50.5 Mb (ftp)(http) TXT
Processed data included within Sample table

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