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在与脐血间充质干细胞共培养过程中,Gata 转录因子家族和 FOG-1 在扩增和分化的脐血源性 CD34 造血干细胞向巨核细胞谱系的上调。

The upregulation of Gata transcription factors family and FOG-1 in expanded and differentiated cord blood-derived CD34 hematopoietic stem cells to megakaryocyte lineage during co-culture with cord blood mesenchymal stem cells.

机构信息

Department of Immunology, Blood Transfusion Research Center, High Institute for Research and Education in Transfusion Medicine (IBTO), Tehran, Iran.

Department of Immunology, Blood Transfusion Research Center, High Institute for Research and Education in Transfusion Medicine (IBTO), Tehran, Iran.

出版信息

Transfus Apher Sci. 2022 Dec;61(6):103481. doi: 10.1016/j.transci.2022.103481. Epub 2022 Jun 6.

Abstract

Umbilical cord blood (UCB) has improved into an attractive and alternative source of allogeneic hematopoietic stem cells (all-HSCs) in clinics and, research for three decades. Recently, it has been shown that the limited cell dose of, this valuable source can be enhanced by the ex vivo expansion of cells in many, ways. We evaluated the expression of the Gata transcription factors family and FOG-1, in expanded and differentiated cord blood-derived CD34 + hematopoietic stem cells to, megakaryocytes lineage., Methods: Separated mononuclear cells were cultured in DMEM complete medium., Harvested cells as a mesenchymal stem cell at 85 % confluency were cultured with, trypsin/EDTA and in 24-well plates. The characteristic analyses of isolated UCB- MSCs, were done by flow cytometry and adipogenic, chondrogenic, and osteogenic, differentiation assays. MACS purified UCB-CD34 + hematopoietic cells cultivated and, differentiated to megakaryocyte progenitor cells in the presence of cytokine cocktail, with UCB-MSCs. Then, the GATA1, GATA2, GATA3, and FOG-1 genes expression, after differentiation to megakaryocyte progenitor cells were performed by quantitative, real-time polymerase chain reaction (PCR)., Results: In this study, the results of real-time-PCR showed that the fold change, expression of GATA-1, FOG-1, and GATA-2 genes after co-culturing with UCB-MSCs, significantly increased to 7.3, 4.7, and 3.3-fold in comparison with control groups;respectively., Conclusion: UCB-MSCs can increase the expansion and differentiation of UCBCD34 + , to megakaryocyte progenitor cells through upregulation of GATA-1, GATA-2, and FOG-1 gene expression.

摘要

脐带血(UCB)在临床和研究中已经成为同种异体造血干细胞(all-HSCs)的有吸引力和替代来源,已经有三十年了。最近,已经表明,这种有价值的来源的有限细胞剂量可以通过多种方法体外扩增细胞来增强。我们评估了 Gata 转录因子家族和 FOG-1 在扩增和分化的脐带血来源 CD34+造血干细胞向巨核细胞谱系中的表达。

方法

分离的单核细胞在 DMEM 完全培养基中培养。在 85%汇合时收获细胞作为间充质干细胞,用胰蛋白酶/EDTA 在 24 孔板中培养。分离的 UCB-MSCs 的特征分析通过流式细胞术和脂肪形成、软骨形成和成骨分化试验进行。MACS 纯化的 UCB-CD34+造血细胞在细胞因子鸡尾酒存在下培养并分化为巨核细胞祖细胞与 UCB-MSCs 共培养。然后,通过定量实时聚合酶链反应(PCR)检测分化为巨核细胞祖细胞后 GATA1、GATA2、GATA3 和 FOG-1 基因的表达。

结果

在这项研究中,实时 PCR 的结果表明,与对照组相比,与 UCB-MSCs 共培养后 GATA-1、FOG-1 和 GATA-2 基因的表达倍数分别显著增加了 7.3、4.7 和 3.3 倍。

结论

UCB-MSCs 可以通过上调 GATA-1、GATA-2 和 FOG-1 基因的表达来增加 UCBCD34+向巨核细胞祖细胞的扩增和分化。

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