T-cell differentiation of multipotent hematopoietic cell line EML in the OP9-DL1 coculture system.
T-cell differentiation of multipotent hematopoietic cell line EML in the OP9-DL1 coculture system.
复制标题
DOI：
10.1016/j.exphem.2009.05.002
复制
发表时间：
2009-08
影响因子：
2.6
通讯作者：
Jurecic, Roland
中科院分区：
文献类型：
--
作者：
Kutlesa, Snjezana;Zayas, Jennifer;Valle, Alexandra;Levy, Robert B.;Jurecic, Roland
多能造血细胞系EML可分化为髓系、红系、巨核系和B淋巴系，但目前尚不清楚EML细胞是否也具有T细胞发育潜能。本研究的目的是确定与表达Notch配体Delta-like 1(OP9-DL1)的OP9基质细胞共培养是否能诱导EML细胞向T细胞分化。EML细胞与对照OP9或OP9-DL1基质细胞在细胞因子(干细胞因子、白介素7和FMS样酪氨酸激酶3配体)的存在下共同培养。通过流式细胞术和逆转录聚合酶链式反应分析细胞表面标志和与T细胞发育特定阶段相关的基因的表达，评估其T细胞谱系的分化。表型、分子和功能分析表明，在有细胞因子的EML/OP9-DL1共培养中，EML细胞系经历了T细胞的定向和分化。在OP9-DL1共培养中，EML细胞已分化为1)类似T细胞发育的双阴性、双阳性和单阳性阶段的细胞；2)启动GATA-3、前T细胞α、RAG-1和T细胞受体-Vβ基因的表达；3)在T细胞受体刺激下产生干扰素-γ。这些结果支持EML细胞系具有T细胞分化能力的观点。值得注意的是，在OP9-DL1共培养中，T系基因的诱导表达和EML细胞向T细胞发育的不同阶段的分化与先前描述的成人造血干细胞和祖细胞的T细胞分化非常相似。因此，EML/OP9-DL1共培养可以作为一个有用的实验系统来研究特定基因在T细胞谱系指定、承诺和分化中的作用。
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. 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. 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-Tα, RAG-1, and T-cell receptor – Vβ genes; and 3) produced interferon-γ in response to T-cell receptor stimulation. 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.
登录
查看更多内容
影响因子：
16
作者：
Harker, N;Naito, T;Kioussis, D
通讯作者：
Kioussis, D
影响因子：
64.8
作者：
Chang, Hannah H.;Hemberg, Martin;Huang, Sui
通讯作者：
Huang, Sui
影响因子：
20.3
作者：
Kirito, K;Fox, N;Kaushansky, K
通讯作者：
Kaushansky, K
影响因子：
30.5
作者：
Allman, D;Sambandam, A;Bhandoola, A
通讯作者：
Bhandoola, A
影响因子：
15.3
作者：
Jaleco, A C;Neves, H;Hooijberg, E;Gameiro, P;Clode, N;Haury, M;Henrique, D;Parreira, L
通讯作者：
Parreira, L