Abstract
To date, different experimental strategies have been developed for the ex vivo 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 ex vivo 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 ex vivo 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.
Highlights
Hematopoiesis is the process of generating mature blood cells and immune cells, which originate in a few populations of hematopoietic stem cells (HSCs) [1, 2]
We focused on the CCL2 gene, which most likely regulates the function of HSCs in the fetal bovine serum (FBS) culture system
CD34+ cells were isolated from human newborn umbilical cord blood (UCB) by the magnetic cell sorting method, and the collected HSCs were transferred into an ex vivo culture system under 2 different conditions
Summary
Hematopoiesis is the process of generating mature blood cells and immune cells, which originate in a few populations of hematopoietic stem cells (HSCs) [1, 2]. HSCs are defined as cells with both the capacity to selfrenew and the ability to differentiate into various types of myeloid and lymphoid lineages [3, 4]. HSCs are frequently used in therapeutic applications to treat hematologic and immune disorders such as leukemia, anemia, congenital immunodeficiencies, metabolic disorders, and autoimmune diseases [5]. Many studies have demonstrated that the CD34+CD38– fraction contains more primitive HSCs that can undergo hematopoiesis than the CD34+CD38+ fraction [7]
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