Neutrophil differentiation from human‐induced pluripotent stem cells

Abstract

Induced pluripotent stem (iPS) cells are of potential value not only for regenerative medicine, but also for disease investigation. The present study describes the development of a neutrophil differentiation system from human iPS cells (hiPSCs) and the analysis of neutrophil function and differentiation. The culture system used consisted of the transfer of hiPSCs onto OP9 cells and their culture with vascular endothelial growth factor (VEGF). After 10 days, TRA 1-85(+) CD34(+) VEGF receptor-2 (VEGFR-2)(high) cells were sorted and co-cultured with OP9 cells in the presence of hematopoietic cytokines for 30 days. Floating cells were collected and subjected to morphological and functional analysis. These hiPSC-derived neutrophils were similar to peripheral blood mature neutrophils in morphology, contained functional neutrophil specific granules, and were equipped with the basic functions such as phagocytosis, superoxide production, and chemotaxis. In the process of differentiation, myeloid cells appeared sequentially from immature myeloblasts to mature segmented neutrophils. Expression patterns of surface antigen, transcription factors, and granule proteins during differentiation were also similar to those of granulopoiesis in normal bone marrow. In conclusion, differentiation of mature neutrophils from hiPSCs was successfully induced in a similar process to normal granulopoiesis using an OP9 co-culture system. This system may be applied to elucidate the pathogenesis of various hematological diseases that affect neutrophils.

Neutrophils alleviate fibrosis in the CCl4‐induced mouse chronic liver injury model

“…Thus, if a sufficient number of neutrophils could be prepared, allogenic neutrophils could be a potential source of cell therapy for cirrhosis. Development of neutrophil differentiation from human‐induced pluripotent stem cells30, 31 would be supportive of such a strategy. Furthermore, delivery of chemoattractive chemokines for neutrophils as well as MMP8 and MMP9 in the liver might be an alternative strategy for the treatment of liver fibrosis.…”

Genetic correction of HAX1 in induced pluripotent stem cells from a patient with severe congenital neutropenia improves defective granulopoiesis

“…Genetic correction of HAX1 in induced pluripotent stem cells from a patient with severe congenital neutropenia improves defective granulopoiesis Masao Kobayashi, 6 Tatsutoshi Nakahata, 2 and Toshio Heike tion system from human iPS cells 17 as well as a serum-and feeder-free monolayer hematopoietic culture system from human ES and iPS cells. 18 In this study, we generate iPS cell lines from an SCN patient with HAX1 gene deficiency and differentiate them into neutrophils in vitro.…”

“…Norio Nakatsuji and Shinya Yamanaka (Kyoto University, Kyoto, Japan), respectively. These human ES and iPS cell lines were maintained on mitomycin-C (Kyowa Hakko Kirin, Tokyo, Japan)-treated SNL feeder cells as described previously 17 and subcultured onto new SNL feeder cells every seven days. …”

Novel Stem Cell–Based Drug Discovery Platforms for Cardiovascular Disease

“…In particular, human iPSCs within various in vitro contexts have been differentiated into endothelium, [29][30][31][32] smooth muscle cells, 30,[33][34][35] monocytes, 36 and neutrophils. [37][38][39] Despite these reports, which describe the emergence of cell populations largely bearing molecular markers of the desired cell type, there is a paucity of functional characterization of the differentiated cell types. There are still significant unanswered questions about the character of the iPSC-derived vascular cellsnamely, whether they faithfully represent the mature phenotype and can perform the many dynamic functions of adult vascular cell types offering a surrogate model system for human primary cells.…”