Abstract

A study was made of the effect that mitomycin C (MitC) treatment of stromal layers of NIH 3T3 cells expressing Jagged1, a ligand of the Notch receptor, exerts on the growth of hematopoietic Lin(–) mouse bone marrow cells in a co-culture system. MitC treatment of stromal cells significantly increased the number of hematopoietic cells and the frequency of colony-forming cells in stromal co-cultures. Transcriptome analysis of control and MitC-treated stromal cell samples was performed by differential RNA sequencing, and genes downregulated by MitC treatment were predominantly associated with the control of cell proliferation, the cell cycle, chromosome segregation, and DNA metabolism. Induction of key hematopoietic cytokines by MitC was not detected by the transcriptome analysis and was therefore not a main factor in the activation of hematopoiesis on the treated stroma. At the same time, the set of the genes most strongly upregulated by MitC treatment is enriched in the genes for cytokines, growth factors, and cell surface proteins, which presumably contribute to enhanced hematopoiesis support on the MitC-treated stroma. Products of some of these genes have been implicated in expansion of hematopoietic stem/progenitor cells in vitro or in vivo.

中文翻译：

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丝裂霉素C处理基质层增强了共培养系统中体外造血的支持

摘要

研究了丝裂霉素C（MitC）处理表达Notch受体配体Jagged1的NIH 3T3细胞的基质层在共培养中对造血Lin（–）小鼠骨髓细胞生长的影响系统。在基质共培养中，基质细胞的MitC处理显着增加了造血细胞的数量和集落形成细胞的频率。对照和经MitC处理的基质细胞样品的转录组分析通过差异RNA测序进行，通过MitC处理下调的基因主要与细胞增殖，细胞周期，染色体分离和DNA代谢的控制相关。转录组分析未检测到由MitC诱导的关键造血细胞因子，因此不是治疗的基质上造血功能激活的主要因素。同时，受MitC处理最强烈上调的一组基因富含细胞因子，生长因子和细胞表面蛋白的基因，这些基因可能有助于增强MitC处理的基质对造血功能的支持。其中一些基因的产物与体外或体内造血干/祖细胞的扩增有关。