Metabolomics,Unknown,Transcriptomics,Genomics,Proteomics

Dataset Information

Gene profiling of platelet-biased long term hematopoietic stem cells

ABSTRACT: Gene profiling of LSKCD150+CD48-CD34- hemetopoietic stem cells expressing (Vwf+) or not expressing a Vwf-EGFP BAC transgene

ORGANISM(S): Mus musculus

SUBMITTER: Nerlov Lab

PROVIDER: E-MEXP-3935 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

Platelet-biased stem cells reside at the apex of the haematopoietic stem-cell hierarchy.

Sanjuan-Pla Alejandra A Macaulay Iain C IC Jensen Christina T CT Woll Petter S PS Luis Tiago C TC Mead Adam A Moore Susan S Carella Cintia C Matsuoka Sahoko S Bouriez Jones Tiphaine T Chowdhury Onima O Stenson Laura L Lutteropp Michael M Green Joanna C A JC Facchini Raffaella R Boukarabila Hanane H Grover Amit A Gambardella Adriana A Thongjuea Supat S Carrelha Joana J Tarrant Paul P Atkinson Deborah D Clark Sally-Ann SA Nerlov Claus C Jacobsen Sten Eirik W SE

Nature 20130811 7470

The blood system is maintained by a small pool of haematopoietic stem cells (HSCs), which are required and sufficient for replenishing all human blood cell lineages at millions of cells per second throughout life. Megakaryocytes in the bone marrow are responsible for the continuous production of platelets in the blood, crucial for preventing bleeding--a common and life-threatening side effect of many cancer therapies--and major efforts are focused at identifying the most suitable cellular and mo ...[more]

PMID: 23934107

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Project description:We discovered that mice that lack Lsd1 in hematopoietic cells were exhibited increased frequencies of CD150+ CD48- lin- c-Kit+ Sca-1+ LT-HSCs, but completely lacked the lin- c-Kit+ Sca-1- myeloid progenitor compartment. To determine the genes altered by Lsd1-loss, CD150+ CD48- lin- c-Kit+ Sca-1+ LT-HSCs from Lsd1fl/fl and Lsd1fl/fl Mx1Cre mice were FACS-purified to be analyzed by gene expression profiling. Primary CD150+ CD48- lin- c-Kit+ Sca-1+ LT-HSCs were isolated by FACS-sorting, from the bone marrow of Lsd1fl/fl and Lsd1fl/fl Mx1Cre animals, one week after the final p(I:C) dose. Total RNA from three biological replicates per genotype was extracted with the RNeasy Micro Kit (QIAGEN), treated with DNaseI, reverse transcribed. RNA extracted from CD150+ CD48- lin- c-Kit+ Sca-1+ cells was amplified with the Ovation Pico WTA RNA Amplification System 2 (NuGEN Technologies). Single-stranded cDNA amplification products were purified using QIAquick PCR Purification Kit (QIAGEN) and labeled with the FL-Ovation cDNA Biotin Module V2 (NuGEN). The Biotin-labeled cDNA was used to hybridize to Affymetrix expression arrays using the Mouse Genome 430 2.0 array platform. Primary CD150+ CD48- lin- c-Kit+ Sca-1+ LT-HSCs were isolated by FACS-sorting, from the bone marrow of Lsd1fl/fl and Lsd1fl/fl Mx1Cre animals, one week after the final p(I:C) dose. Total RNA from three biological replicates per genotype was extracted with the RNeasy Micro Kit (QIAGEN), treated with DNaseI, reverse transcribed. RNA extracted from CD150+ CD48- lin- c-Kit+ Sca-1+ cells was amplified with the Ovation Pico WTA RNA Amplification System 2 (NuGEN Technologies). Single-stranded cDNA amplification products were purified using QIAquick PCR Purification Kit (QIAGEN) and labeled with the FL-Ovation cDNA Biotin Module V2 (NuGEN). The Biotin-labeled cDNA was used to hybridize to Affymetrix expression arrays using the Mouse Genome 430 2.0 array platform. Primary CD150+ CD48- lin- c-Kit+ Sca-1+ LT-HSCs were isolated by FACS-sorting, from the bone marrow of Lsd1fl/fl and Lsd1fl/fl Mx1Cre animals, one week after the final p(I:C) dose. Total RNA from three biological replicates per genotype was extracted with the RNeasy Micro Kit (QIAGEN), treated with DNaseI, reverse transcribed. RNA extracted from CD150+ CD48- lin- c-Kit+ Sca-1+ cells was amplified with the Ovation Pico WTA RNA Amplification System 2 (NuGEN Technologies). Single-stranded cDNA amplification products were purified using QIAquick PCR Purification Kit (QIAGEN) and labeled with the FL-Ovation cDNA Biotin Module V2 (NuGEN). The Biotin-labeled cDNA was used to hybridize to Affymetrix expression arrays using the Mouse Genome 430 2.0 array platform.

Dataset Information

Gene profiling of platelet-biased long term hematopoietic stem cells

Publications

Platelet-biased stem cells reside at the apex of the haematopoietic stem-cell hierarchy.

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