Dataset Information

T-cell differentiation of multipotent hematopoietic cell line EML in the OP9-DL1 coculture system.

ABSTRACT:

Objective

Multipotent hematopoietic cell line EML can differentiate into myeloid, erythroid, megakaryocytic, and B-lymphoid lineages, but it remained unknown whether EML cells have T-cell developmental potential as well. The goal of this study was to determine whether the coculture with OP9 stromal cells expressing Notch ligand Delta-like 1 (OP9-DL1) could induce differentiation of EML cells into T-cell lineage.

Materials and methods

EML cells were cocultured with control OP9 or OP9-DL1 stromal cells in the presence of cytokines (stem cell factor, interleukin-7, and Fms-like tyrosine kinase 3 ligand). Their T-cell lineage differentiation was assessed through flow cytometry and reverse transcription polymerase chain reaction expression analysis of cell surface markers and genes characterizing and associated with specific stages of T-cell development.

Results

The phenotypic, molecular, and functional analysis has revealed that in EML/OP9-DL1 cocultures with cytokines, but not in control EML/OP9 cocultures, EML cell line undergoes T-cell lineage commitment and differentiation. In OP9-DL1 cocultures, EML cell line has differentiated into cells that 1) resembled double-negative, double-positive, and single-positive stages of T-cell development; 2) initiated expression of GATA-3, Pre-Talpha, RAG-1, and T-cell receptor-Vbeta genes; and 3) produced interferon-gamma in response to T-cell receptor stimulation.

Conclusions

These results support the notion that EML cell line has the capacity for T-cell differentiation. Remarkably, induction of T-lineage gene expression and differentiation of EML cells into distinct stages of T-cell development were very similar to previously described T-cell differentiation of adult hematopoietic stem cells and progenitors in OP9-DL1 cocultures. Thus, EML/OP9-DL1 coculture could be a useful experimental system to study the role of particular genes in T-cell lineage specification, commitment, and differentiation.

SUBMITTER: Kutlesa S

PROVIDER: S-EPMC3072798 | biostudies-literature | 2009 Aug

REPOSITORIES: biostudies-literature

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Publications

T-cell differentiation of multipotent hematopoietic cell line EML in the OP9-DL1 coculture system.

Kutlesa Snjezana S Zayas Jennifer J Valle Alexandra A Levy Robert B RB Jurecic Roland R

Experimental hematology 20090515 8

<h4>Objective</h4>Multipotent hematopoietic cell line EML can differentiate into myeloid, erythroid, megakaryocytic, and B-lymphoid lineages, but it remained unknown whether EML cells have T-cell developmental potential as well. The goal of this study was to determine whether the coculture with OP9 stromal cells expressing Notch ligand Delta-like 1 (OP9-DL1) could induce differentiation of EML cells into T-cell lineage.<h4>Materials and methods</h4>EML cells were cocultured with control OP9 or O ...[more]

PMID: 19447159

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Dataset Information

T-cell differentiation of multipotent hematopoietic cell line EML in the OP9-DL1 coculture system.

Objective

Materials and methods

Results

Conclusions

Publications

T-cell differentiation of multipotent hematopoietic cell line EML in the OP9-DL1 coculture system.

Similar Datasets

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