Unknown,Transcriptomics,Genomics,Proteomics

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Gene expression in clonogenic versus non-clonogenic in pooled early passage OCI/AML-4 single cells

ABSTRACT: Identification of genes that regulate clonogenicity of AML cells is hindered by the difficulty of isolating pure populations of cells with defined proliferative abilities. Using early passages of the OCI/AML-4 cell line we sought to determine genes differentially expressed between clonogenic and non-clonogenic cells. We previously shown that OCI/AML-4 clonal siblings display coordinated growth patterns, so that the behaviour of a single cell that is destroyed in the generation of global single cell cDNA may be predicted by the growth patterns of its clonal siblings. Cells were plated in 96 well plates at limiting dilutions. Localized clusters of four cells were identified and micromanipulated such that three of the constituent cells were placed separately into individual microtitre wells containing growth medium and a feeder layer of OP9 cells, while the fourth cell was lysed and processed for global RT-PCR. The cells in each culture well were counted at 2 -3 day intervals until growth stopped. In this manner cDNA was generated from individual clonogenic OCI/AML-4 cells (sibling cells able to generate between 9-100 cells) or from individual non-clonogenic OCI/AML-4 cells (sibling cells were not able to generate more than 8 cells). Respective single cell cDNA was then pooled and compared by hybridization to cDNA microarrays. Using early passages of the OCI/AML-4 cell line cDNA was obtained from 17 clonogenic single cells (sibling cells able to generate between 9-100 cells) and 20 non-clonogenic single cells (sibling cells were not able to generate more than 8 cells). These cDNAs were pooled and hybridized to the cDNA microarray was conducted as three replicates, including one replicate which was a dye-swap.

ORGANISM(S): Homo sapiens

SUBMITTER: Christine Ichim

PROVIDER: E-GEOD-28612 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

Identification of a role for the nuclear receptor EAR-2 in the maintenance of clonogenic status within the leukemia cell hierarchy.

Ichim C V CV Atkins H L HL Iscove N N NN Wells R A RA

Leukemia 20110603 11

Identification of genes that regulate clonogenicity of acute myelogenous leukemia (AML) cells is hindered by the difficulty of isolating pure populations of cells with defined proliferative abilities. By analyzing the growth of clonal siblings in low passage cultures of the cell line OCI/AML4 we resolved this heterogeneous population into strata of distinct clonogenic potential, permitting analysis of the transcriptional signature of single cells with defined proliferative abilities. By microarr ...[more]

PMID: 21637284

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Dataset Information

Gene expression in clonogenic versus non-clonogenic in pooled early passage OCI/AML-4 single cells

Publications

Identification of a role for the nuclear receptor EAR-2 in the maintenance of clonogenic status within the leukemia cell hierarchy.

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