Differentiation of Human Embryonic and Induced Pluripotent Stem Cells into Blood Cells in Coculture with Murine Stromal Cells

Ma, Feng; Gu, Yanzheng; Nishihama, Natsumi; Yang, Wenyu; Ebihara, Yasuhiro; Tsuji, Kohichiro

doi:10.1007/978-1-61779-267-0_23

Cited by 5 publications

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“…We examined the hematopoietic differentiation ability of EMS-iPS cells in comparison to that of control iPS cells, which were generated from healthy donors using common procedures. To generate hematopoietic cells from EMS-iPS cells and control iPS cells, a co-culture system with AGM-S3 cells was used, as previously reported [ 23 ] [ 21 ]. When colonies of undifferentiated EMS-iPS cells or control iPS cells were co-cultured with AGM-S3 cells, cells with a cobblestone morphology were detected at the peripheries of the colonies at day 11 or 12 ( Fig.…”

Section: Resultsmentioning

confidence: 99%

“…iPS cells were maintained and passaged weekly on mitomycin C-treated MEF feeder cells, as described previously [ 17 ] [ 19 ] [ 21 ]. To evaluate the potential of iPS cells to differentiate into hematopoietic cells, iPS cells were co-cultured with the murine stromal cell line, AGM-S3 [ 17 ] [ 22 ] [ 23 ]. Briefly, 50 undifferentiated iPS cell colonies (each consisting of approximately 1 × 10 3 cells) were transferred to each well of a 6-well plate (Sumitomo Bakelite Co), which contained confluent 15 Gy-irradiated AGM-S3 cells (approximately 2–3 × 10 5 cells per well).…”

Section: Methodsmentioning

confidence: 99%

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Screening of Drugs to Treat 8p11 Myeloproliferative Syndrome Using Patient-Derived Induced Pluripotent Stem Cells with Fusion Gene CEP110-FGFR1

et al. 2015

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Induced pluripotent stem (iPS) cells provide powerful tools for studying disease mechanisms and developing therapies for diseases. The 8p11 myeloproliferative syndrome (EMS) is an aggressive chronic myeloproliferative disorder (MPD) that is caused by constitutive activation of fibroblast growth factor receptor 1. EMS is rare and, consequently, effective treatment for this disease has not been established. Here, iPS cells were generated from an EMS patient (EMS-iPS cells) to assist the development of effective therapies for EMS. When iPS cells were co-cultured with murine embryonic stromal cells, EMS-iPS cells produced more hematopoietic progenitor and hematopoietic cells, and CD34+ cells derived from EMS-iPS cells exhibited 3.2–7.2-fold more macrophage and erythroid colony forming units (CFUs) than those derived from control iPS cells. These data indicate that EMS-iPS cells have an increased hematopoietic differentiation capacity, which is characteristic of MPDs. To determine whether a tyrosine kinase inhibitor (TKI) could suppress the increased number of CFUs formed by EMS-iPS-induced CD34+ cells, cells were treated with one of four TKIs (CHIR258, PKC 412, ponatinib, and imatinib). CHIR258, PKC 412, and ponatinib reduced the number of CFUs formed by EMS-iPS-induced CD34+ cells in a dose-dependent manner, whereas imatinib did not. Similar effects were observed on primary peripheral blood cells (more than 90% of which were blasts) isolated from the patient. This study provides evidence that the EMS-iPS cell line is a useful tool for the screening of drugs to treat EMS and to investigate the mechanism underlying this disease.

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Section: Resultsmentioning

confidence: 99%