Department of Chemical Engineering, National Cheng Kung University, Tainan 701, Taiwan.
Department of Chemical Engineering and Materials Science, Yuan Ze University, Chung-Li, Taoyuan City 320, Taiwan.
Aging (Albany NY). 2021 Aug 10;13(15):19878-19893. doi: 10.18632/aging.203398.
To date, different experimental strategies have been developed for the expansion of human hematopoietic stem cells (HSCs) for clinical applications. However, differences in the genomic function of expanded HSCs under different culture systems remain unclear. In this study, we compared the gene expression profiles of HSCs in expanded serum (10% FBS, fetal bovine serum) and serum-free culture systems and analyzed the molecular functions of differentially expressed genes using microarray chips. We identified 839 differentially expressed genes between the two culture systems. These genes were enriched in the TNF -regulated inflammatory pathway in an FBS culture system. In addition, the mRNA expression of CCL2 (C-C motif chemokine ligand 2), TNF (tumor necrosis factor) and FOS (FBJ murine osteosarcoma viral oncogene homolog) was validated by RT-qPCR. Our data revealed that expansion of HSCs using the FBS culture system induces an inflammatory response and high CD38 expression, indicating that this system might activate an inflammatory pathway and induce expression of the cancer marker CD38 during ex vivo expansion of HSCs. This study provides a transcriptional profile and new insights into the genomic functions of HSCs under different expanded cultures.
迄今为止，已经开发出了多种不同的实验策略来扩增人类造血干细胞（HSCs），以应用于临床。然而，不同培养体系下扩增的 HSCs 的基因组功能差异尚不清楚。在本研究中，我们比较了在扩增血清（10% FBS，胎牛血清）和无血清培养体系中 HSCs 的基因表达谱，并使用微阵列芯片分析了差异表达基因的分子功能。我们在两种培养体系中鉴定出了 839 个差异表达基因。这些基因在 FBS 培养体系中富集于 TNF 调控的炎症通路。此外，我们通过 RT-qPCR 验证了 CCL2（C-C 基序趋化因子配体 2）、TNF（肿瘤坏死因子）和 FOS（FBJ 鼠骨肉瘤病毒癌基因同源物）的 mRNA 表达。我们的数据表明，使用 FBS 培养体系扩增 HSCs 会引发炎症反应和高 CD38 表达，这表明该体系可能在 HSCs 的体外扩增过程中激活炎症通路并诱导癌症标志物 CD38 的表达。本研究提供了一个转录谱，并为不同扩增培养体系下 HSCs 的基因组功能提供了新的见解。
