Journal: Stem Cell Research & Therapy

Article Title: Ricolinostat promotes the generation of megakaryocyte progenitors from human hematopoietic stem and progenitor cells

doi: 10.1186/s13287-022-02722-5

Figure Lengend Snippet: Induction of MkPs into MKs. A Schematic representation of the stepwise protocol for HSPC differentiation into MKs. CD34 + HSPCs were cultured for 7 days in the presence of SCF, TPO, IL-3, IL-6, and FLT3-L for HSPC expansion (stage I). Cells obtained from the first stage were cultured for 7 days in the presence of SCF, TPO, IL-3, IL-6, and IL-11, with or without Ricolinostat for megakaryocytic lineage specification (stage II). Megakaryocytic lineage cells obtained from the second stage were cultured without Ricolinostat for 7–14 days in the presence of cytokines for MK maturation and platelet production (stage III). B Flow cytometry analysis of CD41a and CD42b expression in cells obtained from the third stage on day 21. C Percentage of CD41a + CD42b + cells on day 21. D Number of CD41a + CD42b + cells from per 10 5 HSPCs on day 21. E Identification of morphology and structure of MKs by Wright's Giemsa staining. Scale bars, 10 μm. F Immunofluorescence analysis showed the expression and location of MK-specific markers CD41, CD42b, α-Tubulin, vWF, PF4, and THBS-1 in cells on day 21. Scale bars, 5 μm. G , H Expression of THBS-1 in the culture supernatant and cells on day 21 measured by ELISA. I , J Expression of PF4 in the culture supernatant and cells on day 21 measured by ELISA. Results are expressed as the mean ± SEM from three independent experiments. Unpaired Student’s t -test, * p < 0.05

Article Snippet: To detect and quantify human MK progenitors, on day 7 of stage II, 40,000 cells were plated in a serum-free collagen-based system using the MegaCult™-C Complete Kit with Cytokines (STEMCELL Technologies) according to the manufacturer’s instructions [ ].

Techniques: Cell Culture, Flow Cytometry, Expressing, Staining, Immunofluorescence, Enzyme-linked Immunosorbent Assay