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Fig. 3 | Stem Cell Research & Therapy

Fig. 3

From: Thrombopoietin knock-in augments platelet generation from human embryonic stem cells

Fig. 3

TPO knock-in augments megakaryocytic differentiation. a Schematic of megakaryocytic differentiation from HPCs to MKs and PLTs using mAGM-S3 stromal cell coculture. b Representative cell morphology at day 6 of megakaryocytic differentiation from Ctr or TPO-KI-1/-2 H1 hESCs. Scale bar = 40 μm. Large cells indicated by white arrows. c Distribution of sizes of megakaryocytes formed at day 6 in (a) as measured with microscopy. Data shown as mean ± SEM (n = 3). *P < 0.05. d Ploidy distribution of megakaryocytes analyzed by staining cellular DNA with propidium iodide (PI). Data shown as mean ± SEM (n = 3). *P < 0.05; **P < 0.01. e, f Flow cytometer analysis for percentage of CD41a+ (e) or CD41a+CD42b+ (f) megakaryocytes at day 3 or day 6 of MK differentiation, respectively. Data shown as mean ± SEM (n = 3). *P < 0.05; **P < 0.01. g qRT-PCR analysis of megakaryocytic-associated markers (GATA1, FLI-1, RUNX1, FOG-1, NF-E2, ITGB3) in Ctr or TPO-KI-1/-2 H1 hESCs for indicated times. ACTIN used as internal control. All values normalized to level (=1) of mRNA in Ctr at day 0 of MK differentiation. Data shown as mean ± SEM (n = 3). *P < 0.05; **P < 0.01; NS, not significant. Ctr control, HPC hematopoietic progenitor cell, IL interleukin, MK megakaryocyte, PLT platelet, TPO thrombopoietin

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